Loading...
Browse
Search Results
ThesisItem Open Access PHYSICO-CHEMICAL, FUNCTIONAL AND THERMAL CHARACTERIZATION OF DIFFERENTLY TREATED BARNYARD (Echinochloa esculenta), FOXTAIL (Setaria italica) AND LITTLE (Panicum sumatrense) MILLET FLOURS(PROFESSOR JAYASHANKAR TELANGANA STATE AGRICULTURAL UNIVERSITY, 2022-03-01) SHREEJA, K; HYMAVATHI, T.V.Today's health trends shift from reactive to proactive and from treatment to prevention. There is a growing awareness of the importance of health and nutrition and a growing desire to lead an active and healthy lifestyle. This is driving the consumption of millet with increasing awareness of its health benefits. Epidemiological studies have shown that regular consumption of millets reduces the risk of heart disease, minimizes the onset of type II diabetes, improves the digestive system, lowers the risk of cancer, detoxifies the body, and increases immunity including respiratory health and also Parkinson’s disease. Several initiatives are also being undertaken toward enhancing millet cultivation and consumption to reduce health risks. To improve the consumption of millets to a desirable level, in addition to sufficient production, products of commercial importance should be made available in the market. Flour is a basic ingredient for many products across the world. Earlier studies on pretreated flours were not fully investigated. Thus, the aim of the study was to elucidate the differently treated millet flours in terms of physical, functional, thermal, pasting, rheological, nutritional and antioxidant properties. In the present study, three types of minor millets viz., barnyard (BM), foxtail (FM) xxiii and little millet (LM) were selected. Both the whole and dehulled grains were subjected to soaking, germination, fermentation, dry heating, hydrothermal and extrusion treatments after optimization and converted into flours. The resultant flours were examined for physical, functional, pasting, nutritional and antioxidant properties. Storage stability was studied for 90 days. Overall, there was a significant effect of the treatments in all the grains in the different properties studied. Multivariate analysis revealed that the effect was due to treatments than the grains. The mean values of loose bulk density (LBD), tapped bulk density (TBD), color attributes L*, a*, b*, pH, and sedimentation value (SV) were 0.54 (g/ml), 0.68 (g/ml), 18.37 27.88, 3.20, 6.57 and 1.92ml, respectively. LBD of the whole, whole soaked (WS), germinated, fermented and whole hydrothermal (WHT) flours were lower than their respective means. WS, WHT, whole extruded, fermented, and dry heated flours have lower TBD than their respective means. Evaluation of functional properties indicated that water holding capacity (WHC), water absorption capacity (WAC), water solubility index (WSI), swelling power (SP), and swelling index (SI) were significantly improved by extrusion, hydrothermal treatment, germination, and dry heating. The mean WHC, WAC, WSI, SP, SI, foaming capacity (FC) and oil retention capacity (ORC) were 181.44 (%), 6.61%, 3.89, 318.19, 1.41 (g/dl), 13.42 % and 2.14%, respectively. Apart from untreated, whole soaked and whole dried (WDr) flours exhibited higher foaming capacity (FC%) followed by their dehulled counterparts. Foaming was not observed in extruded flours. Whole extruded (WE) and dehulled soaked (DS) flours exhibited highest and lowest ORC, respectively. Measurements of gelatinization temperature revealed a significant effect of treatments and grain on the gelatinization temperature of the millet flours. There was no significant effect of grains on Tc and enthalpy. Hydrothermally treated (HT) followed by, extruded and soaked exhibited the lowest onset, peak and conclusion temperature, while the dehulled fermented flours (DFF) were highest. Enthalpy was lowest in germinated flours (GFFs) and highest in dehulled dry heated flours (DDrF) (P< 0.05). Extrusion significantly improved the mean pasting viscosity (PV) from 244.80 to 4034.33 cP in whole and 227.87 to 3606.33 cP in dehulled flours followed by germinated flours (244.80 to 547.53). DSF, DDrF, DHTF, DFF, and dehulled flours exhibited similar (P<0.05) high peak viscosity (PKV). Extruded followed by germinated flours have very low PKV. Break down viscosity (BV) was significantly high in DDrF, DFF at par with dehulled untreated flours. DSF followed by WHTF and DHTF followed by DFF showed xxiv very high holding viscosity (HV) and Final viscosity (FV) than others, while those of germinated and extruded flours were lower than others. Setback viscosity (SV) of extruded flours, DFF, WHTF and DHTF was significantly higher than others, with no difference among them. Storage and loss modulus were increased in all the treated whole barnyard flour doughs except extruded dough. Among all the millet flours, extruded flours exhibited the lowest G′′ and G′. Significantly higher ash, protein, and crude fibre content was in WEF, DHTF and DDrF, respectively. No significant effect of treatments on carbohydrate and fat content. More remarkable improvement was in whole grain treatments than that of dehulled grain. Zinc (6.93 mg/100 g), iron (4.93 mg/100 g), and calcium (42.88 mg/100 g) contents were enhanced in DSF, DSF and Germinated flours, respectively (p<0.05), while extrusion improved potassium levels. Dry heating lowered the protein content. HTF and WFF increased TFC, while that of TPC and tannins increased in extruded flours, which was reflected in lower IC50 and increased DPPH % inhibition values. Dehulled treated flours reduced a higher percentage of oxalates (78.2 to 128.5%) than whole treated (50.6 to 72.2%). Storage of all the flours did not adversely effect the shelf life. Thus, the study demonstrated that treated flours can be developed with enhanced nutrition, physical, functional, thermal, and rheological properties. Further studies on the in vitro digestibility and glycemic index would help promote these flours as functional ingredientThesisItem Open Access TRANSCRIPTOME PROFILING AND ANALYSIS OF Fe AND Zn RESPONSIVE GENES IN PEARL MILLET (Pennisetum glaucum L.)(PROFESSOR JAYASHANKAR TELANGANA STATE AGRICULTURAL UNIVERSITY, 2022-02-07) ANJALI, C; VANISRI, SMicronutrient malnutrition because of Fe and Zn deficiencies is posing a serious health issue in developing countries worldwide. To combat the micronutrient-based hidden hunger, biofortification is an efficient and cost-effective method to enhance the nutrient contents of crops by breeding techniques. Spearheaded by the HarvestPlus Program of Consultative Group for International Agricultural Research (CGIAR), global crop biofortification research was initiated and focused primarily on seven major staple crops including pearl millet, targeting three important micronutrients (Fe, Zn, and vitamin A). Pearl millet (Pennisetum glaucum L.) is a staple for millions of families in dryland tropics and reported varying concentrations of Fe and Zn in germplasm. Studies on the functional characterization of differentially expressed genes (DEGs) and their dynamic role in Fe and Zn pathways will accelerate the biofortified cultivar development. Transcriptome profiling of leaf and root samples of a pearl millet inbred ICMB 1505 under Fe and Zn stress conditions were carried out wherein ten to twelve days old seedlings were exposed to Fe and Zn stress treatments (–Fe–Zn, –Fe+Zn, +Fe–Zn, and +Fe+Zn) for 12 days. Total RNA was extracted from the treated samples followed by cDNA synthesis, cDNA library preparation, and sequencing of the constructed cDNA libraries. A total of 37,093 DEGs under different combinations of leaf and root stress conditions were identified, of which, 7023 and 9996 DEGs were reported in leaf and root stress conditions, respectively. Among the 11,429 unique DEGs, 8605 were annotated to cellular, biological, molecular functions and 458 DEGs were assigned to 39 pathways. The results revealed the expression of major genes related to mugineic acid pathway, phytohormones, chlorophyll biosynthesis, photosynthesis, and carbohydrate metabolism during Fe and Zn starvation in pearl millet. The analysis reported the expression of nicotianamine synthase (NAS), S-adenosyl L- methionine (SAM) synthetase, 2'-deoxymugineic acid (DMA), 2'- deoxymugineic- acid 2'-dioxygenase (IDS3), FER-like transcription factor genes involved in uptake mechanism and oligopeptide transporter 3 (OPT3), Zn-transporter 3 & 5, Zinc induced facilitator 1 (ZIF1), heavy metal ATPases (HMA) genes for transport of Fe and Zn. The study also discussed the expressional changes of several cellular pathway genes and their regulation in Fe and Zn homeostasis under their deficient conditions. The cross-talks between the Fe and Zn provided information on their dual and opposite regulation of key uptake and transporter genes under deficiency conditions. The identified Fe and Zn homeostasis-related genes from the transcriptomic data were annotated onto the pearl millet genome wherein a high number of genes were distributed on chromosome 3 and a low number of genes were distributed on chromosome 7. The orthologues for the top 42 genes were identified in Oryza sativa, Zea mays and Sorghum bicolor along with their annotations and chromosomal positions in respective crops. Moreover, the orthologues of the uncharacterized genes among the top 42 genes, selected based on gene ontology (GO) terms (involved in Fe and Zn homeostasis) reported being major Fe and Zn uptake (NAS, DMAS) and transporter genes (zinc transporter 4 and 9, YSL transporters) in rice, maize, and sorghum. The orthologues identified in rice, maize, and sorghum aids in the genetic biofortification of nutrient contents in these crops. The gene structures of the identified orthologues were represented which helps in understanding the exon-intron positions and evolutionary changes among the species. The knowledge of the position of the above-identified genes in pearl millet, rice, maize, and sorghum crops can be utilized in the genetic biofortification of these crops for Fe and Zn contents by genetic engineering and other breeding programs. Our results assist in developing Fe and Zn-efficient pearl millet varieties through development of genic-SNPs for Fe and Zn responsive genes and their utilization in an ongoing biofortification breeding program to ameliorate malnutrition in the dryland tropics of South Asia and Sub-Saharan Africa.ThesisItem Open Access TRANSCRIPTOME PROFILING AND ANALYSIS OF Fe AND Zn RESPONSIVE GENES IN PEARL MILLET (Pennisetum glaucum L.)(PROFESSOR JAYASHANKAR TELANGANA STATE AGRICULTURAL UNIVERSITY, 2022-02-07) ANJALI, C; VANISRI, SMicronutrient malnutrition because of Fe and Zn deficiencies is posing a serious health issue in developing countries worldwide. To combat the micronutrient-based hidden hunger, biofortification is an efficient and cost-effective method to enhance the nutrient contents of crops by breeding techniques. Spearheaded by the HarvestPlus Program of Consultative Group for International Agricultural Research (CGIAR), global crop biofortification research was initiated and focused primarily on seven major staple crops including pearl millet, targeting three important micronutrients (Fe, Zn, and vitamin A). Pearl millet (Pennisetum glaucum L.) is a staple for millions of families in dryland tropics and reported varying concentrations of Fe and Zn in germplasm. Studies on the functional characterization of differentially expressed genes (DEGs) and their dynamic role in Fe and Zn pathways will accelerate the biofortified cultivar development. Transcriptome profiling of leaf and root samples of a pearl millet inbred ICMB 1505 under Fe and Zn stress conditions were carried out wherein ten to twelve days old seedlings were exposed to Fe and Zn stress treatments (–Fe–Zn, –Fe+Zn, +Fe–Zn, and +Fe+Zn) for 12 days. Total RNA was extracted from the treated samples followed by cDNA synthesis, cDNA library preparation, and sequencing of the constructed cDNA libraries. A total of 37,093 DEGs under different combinations of leaf and root stress conditions were identified, of which, 7023 and 9996 DEGs were reported in leaf and root stress conditions, respectively. Among the 11,429 unique DEGs, 8605 were annotated to cellular, biological, molecular functions and 458 DEGs were assigned to 39 pathways. The results revealed the expression of major genes related to mugineic acid pathway, phytohormones, chlorophyll biosynthesis, photosynthesis, and carbohydrate metabolism during Fe and Zn starvation in pearl millet. The analysis reported the expression of nicotianamine synthase (NAS), S-adenosyl L- methionine (SAM) synthetase, 2'-deoxymugineic acid (DMA), 2'- deoxymugineic- acid 2'-dioxygenase (IDS3), FER-like transcription factor genes involved in uptake mechanism and oligopeptide transporter 3 (OPT3), Zn-transporter 3 & 5, Zinc induced facilitator 1 (ZIF1), heavy metal ATPases (HMA) genes for transport of Fe and Zn. The study also discussed the expressional changes of several cellular pathway genes and their regulation in Fe and Zn homeostasis under their deficient conditions. The cross-talks between the Fe and Zn provided information on their dual and opposite regulation of key uptake and transporter genes under deficiency conditions. The identified Fe and Zn homeostasis-related genes from the transcriptomic data were annotated onto the pearl millet genome wherein a high number of genes were distributed on chromosome 3 and a low number of genes were distributed on chromosome 7. The orthologues for the top 42 genes were identified in Oryza sativa, Zea mays and Sorghum bicolor along with their annotations and chromosomal positions in respective crops. Moreover, the orthologues of the uncharacterized genes among the top 42 genes, selected based on gene ontology (GO) terms (involved in Fe and Zn homeostasis) reported being major Fe and Zn uptake (NAS, DMAS) and transporter genes (zinc transporter 4 and 9, YSL transporters) in rice, maize, and sorghum. The orthologues identified in rice, maize, and sorghum aids in the genetic biofortification of nutrient contents in these crops. The gene structures of the identified orthologues were represented which helps in understanding the exon-intron positions and evolutionary changes among the species. The knowledge of the position of the above-identified genes in pearl millet, rice, maize, and sorghum crops can be utilized in the genetic biofortification of these crops for Fe and Zn contents by genetic engineering and other breeding programs. Our results assist in developing Fe and Zn-efficient pearl millet varieties through development of genic-SNPs for Fe and Zn responsive genes and their utilization in an ongoing biofortification breeding program to ameliorate malnutrition in the dryland tropics of South Asia and Sub-Saharan Africa. Micronutrient malnutrition because of Fe and Zn deficiencies is posing a serious health issue in developing countries worldwide. To combat the micronutrient-based hidden hunger, biofortification is an efficient and cost-effective method to enhance the nutrient contents of crops by breeding techniques. Spearheaded by the HarvestPlus Program of Consultative Group for International Agricultural Research (CGIAR), global crop biofortification research was initiated and focused primarily on seven major staple crops including pearl millet, targeting three important micronutrients (Fe, Zn, and vitamin A). Pearl millet (Pennisetum glaucum L.) is a staple for millions of families in dryland tropics and reported varying concentrations of Fe and Zn in germplasm. Studies on the functional characterization of differentially expressed genes (DEGs) and their dynamic role in Fe and Zn pathways will accelerate the biofortified cultivar development. Transcriptome profiling of leaf and root samples of a pearl millet inbred ICMB 1505 under Fe and Zn stress conditions were carried out wherein ten to twelve days old seedlings were exposed to Fe and Zn stress treatments (–Fe–Zn, –Fe+Zn, +Fe–Zn, and +Fe+Zn) for 12 days. Total RNA was extracted from the treated samples followed by cDNA synthesis, cDNA library preparation, and sequencing of the constructed cDNA libraries. A total of 37,093 DEGs under different combinations of leaf and root stress conditions were identified, of which, 7023 and 9996 DEGs were reported in leaf and root stress conditions, respectively. Among the 11,429 unique DEGs, 8605 were annotated to cellular, biological, molecular functions and 458 DEGs were assigned to 39 pathways. The results revealed the expression of major genes related to mugineic acid pathway, phytohormones, chlorophyll biosynthesis, photosynthesis, and carbohydrate metabolism during Fe and Zn starvation in pearl millet. The analysis reported the expression of nicotianamine synthase (NAS), S-adenosyl L- methionine (SAM) synthetase, 2'-deoxymugineic acid (DMA), 2'- deoxymugineic- acid 2'-dioxygenase (IDS3), FER-like transcription factor genes involved in uptake mechanism and oligopeptide transporter 3 (OPT3), Zn-transporter 3 & 5, Zinc induced facilitator 1 (ZIF1), heavy metal ATPases (HMA) genes for transport of Fe and Zn. The study also discussed the expressional changes of several cellular pathway genes and their regulation in Fe and Zn homeostasis under their deficient conditions. The cross-talks between the Fe and Zn provided information on their dual and opposite regulation of key uptake and transporter genes under deficiency conditions. The identified Fe and Zn homeostasis-related genes from the transcriptomic data were annotated onto the pearl millet genome wherein a high number of genes were distributed on chromosome 3 and a low number of genes were distributed on chromosome 7. The orthologues for the top 42 genes were identified in Oryza sativa, Zea mays and Sorghum bicolor along with their annotations and chromosomal positions in respective crops. Moreover, the orthologues of the uncharacterized genes among the top 42 genes, selected based on gene ontology (GO) terms (involved in Fe and Zn homeostasis) reported being major Fe and Zn uptake (NAS, DMAS) and transporter genes (zinc transporter 4 and 9, YSL transporters) in rice, maize, and sorghum. The orthologues identified in rice, maize, and sorghum aids in the genetic biofortification of nutrient contents in these crops. The gene structures of the identified orthologues were represented which helps in understanding the exon-intron positions and evolutionary changes among the species. The knowledge of the position of the above-identified genes in pearl millet, rice, maize, and sorghum crops can be utilized in the genetic biofortification of these crops for Fe and Zn contents by genetic engineering and other breeding programs. Our results assist in developing Fe and Zn-efficient pearl millet varieties through development of genic-SNPs for Fe and Zn responsive genes and their utilization in an ongoing biofortification breeding program to ameliorate malnutrition in the dryland tropics of South Asia and Sub-Saharan Africa.ThesisItem Open Access GENETIC ANALYSIS FOR YIELD AND SHOOT FLY RESISTANCE IN CMS BASED HYBRIDS OF SORGHUM [Sorghum bicolor (L.) Moench(PROFESSOR JAYASHANKAR TELANGANA STATE AGRICULTURAL UNIVERSITY, 2022-02-07) SAI KIRAN, V; SHIVANI, DThe present investigation was conducted to study the performance of the 48 hybrids and their parents with respect to grain yield and shoot fly resistance in sorghum. The experiment was conducted using 14 parental lines, which included six females, viz., ICSA 418, ICSA 419, ICSA 427, ICSA 433, ICSA 435 and ICSA 29004 (male sterile, A lines) and eight male lines, ICSR 13004, ICSR 13009, ICSR 13025, ICSR 13031, ICSR 13042, ICSR 13043, ICSR 13046 and ICSR 29 (Restorers, R lines and high yielding) differed in levels of resistance against sorghum shoot fly. The 48 experimental hybrids developed using these A lines and R lines were evaluated at three locations: viz., Regional Agricultural Research Station (RARS), Palem; Agricultural Research Station (ARS), Tandur and College Farm, Rajendranagar. Four standard checks were used in the investigation, and out of the four checks, two (CSV 41 and CSH 16) were used for yield analysis and two (IS 18551 as resistant and Swarna as susceptible) for shoot fly screening. In yield analysis experiment, each plot consisted of 2 rows of 4 metres each, with an inter row spacing of 45 cm and an intra row spacing of 10 cm. Normal recommended cultural practices were followed during the evaluation to raise a successful crop for better phenotypic expression of characters. In each location, five competitive plants were randomly selected from each plot for recording observations on days to 50 % flowering, days to maturity, plant height, 100 grain weight (g), grain yield per plant (g), grain yield per plot (kg), fodder yield per plant (g) and fodder yield per plot (kg). Days to 50 % flowering and days to maturity were recorded on a plot basis. In the shoot fly screening experiment, all the protection measures were avoided. Data from all the component characters associated with shoot fly resistance, such as seedling vigour, seedling glossiness, number of eggs per plant (14 DAE), trichome density on upper and lower leaf surfaces, and deadhearts per cent (21 DAE and 28 DAE) were collected from each genotype. All the data from both experiments was analyzed using windostat software (version 9.1) from indostat services at the University Computer Centre, Rajendranagar, Hyderabad. In addition to the field screening of genotypes for shoot fly resistance, a molecular screening experiment was carried out using 20 reported markers linked to shoot fly resistance to check whether the markers so far reported to be linked to shoot fly resistance can be used for differentiating the resistance and susceptibility among the parental genotypes (six lines and eight testers) used in the present study. The results from the investigation revealed that good combiners for grain yield per plant need to be desirable combiners for lower shoot fly deadhearts per cent and vice versa. The general combining ability of parents for grain yield per plant and shoot fly deadhearts per cent (28 DAE) showed that three parents involving one line and two testers viz., ICSA 433, ICSR 13009 and ICSR 13025, respectively, were found to be good general combiners for both grain yield per plant as well as lower shoot fly deadhearts per cent. Most of the good specific combiners for grain yield per plant were not desirable combiners for deadhearts per cent and vice versa. About 8 hybrids viz., ICSA 419 × ICSR 13043 (2.419 and -8.108), ICSA 427 × ICSR 13004 (33.394 and - 6.398), ICSA 427 × ICSR 13031(2.625 and -4.226), ICSA 433 × ICSR 13043 (30.482 and -5.741), ICSA 29004 × ICSR 13025 (14.130 and -4.027) and ICSA 29004 × ICSR 13046 (2.536 and -10.272) were found to be the best specific combiners for both grain yield per plant and lower shoot fly deadhearts per cent. It was found from the investigation that hybrids with high grain yield per plant need not to be with low deadhearts per cent score and vice versa. About five hybrids (displayed in the order of merit), viz., ICSA 435 × ICSR 13042 (95.285 g), ICSA 427 × ICSR 13031 (92.868 g), ICSA 419 × ICSR 13042 (84.910 g), ICSA 435 × ICSR 13031 (84.452 g) and ICSA 418 × ICSR 13009 (83.165 g) recorded high grain yield per plant as well as resistance (damage score of 3.0) against sorghum shoot fly whereas the high yielding hybrid viz., ICSA 427 × ICSR 13046 (85.107), was susceptible to shoot fly damage with high per cent of deadhearts (damage score of 7.0). Even the resistant hybrids (damage score of 3.0) viz., ICSA 419 × ICSR 13009, ICSA 427 × ICSR 13009, ICSA 433 × ICSR 13004, ICSA 433 × ICSR 13025 and ICSA 433 × ICSR 13043 were found to be low yielders across environments. Therefore, it was suggested from the present investigation that hybrids viz., ICSA 435 × ICSR 13042, ICSA 427 × ICSR 13031, ICSA 419 × ICSR 13042, ICSA 435 × ICSR 13031 and ICSA 418 × ICSR 13009 had both high yielding and shoot fly resistance characteristics and could be commercially exploited after sufficient testing. The stability parameters for component traits revealed that as many as 15 and 11 genotypes turned out to be stable for days to 50 % flowering and days to maturity, respectively, with lower means than parents and hybrids. Similarly, for plant height 12, for 100 Grain weight 11, for grain yield per plot 14, for grain yield per plant 23, for fodder yield per plot 15 and for fodder yield per plant 23, were found to be stable, i.e. they had non-significant bi and S2 di values. Female ICSA 427 and ICSA 433 recorded high grain yield per plant with better gca effects and stability for grain yield per plant and yield contributing characters. The male parent, 13031 had high grain yield along with good general combining ability and stability of performance for grain yield per plant. Three hybrids, viz., ICSA 418 × ICSR 13025, ICSA 29004 × ICSR 29 and ICSA 419 × ICSR 29 had high mean performance with average stability, high standard heterosis performance (over CSV 41 or CSH 16 or both) and desirable sca effects for grain yield and some of its component traits. Most of the hybrids recorded high grain yield per plant however, the stability performance of those hybrids was not satisfactory. Interestingly, from the entire investigation, three hybrids viz., ICSA 418 × ICSR 13025 (112.883 g), ICSA 419 × ICSR 29 (106.082 g) and ICSA 29004 × ICSR 29 (93.710 g) were found to be highly stable grain yielders. However, they showed moderate resistance against sorghum shoot fly. Therefore, these hybrids would be used for further screening of different locations. From the molecular screening of parents, it was found that among the 20 SSR markers used; only one marker viz., Xnhsbm 1011 showed differentiation of resistance for sorghum shoot fly among the parental genotypes screened. The marker was polymorphic between the lines and testers (showing a difference in banding pattern). Morphological screening studies revealed that, the performance against shoot fly was different among the parental genotypes. Lines were found to be moderately susceptible to susceptible against shoot fly at phenotypic level, whereas all eight testers were moderately resistant against shoot fly. These parents could be differentiated by this marker (Xnhsbm 1011) which showed an allele size of 150 bp in all the lines (moderately susceptible) and 160 bp in all the testers (moderately resistant). From the investigation, it can be concluded that both additive and non-additive gene effects are important, with a predominance of non-additive gene effects in governing the traits associated with yield and shoot fly resistance in sorghum. Crossing between the resistant or moderately resistant lines endowed with different resistant mechanisms is likely to produce stable lines with desirable traits. Hybrids identified in the present investigation, viz., ICSA 435 × ICSR 13042, ICSA 427 × ICSR 13031, ICSA 419 × ICSR 13042, ICSA 435 × ICSR 13031 and ICSA 418 × ICSR 13009, having superior grain yield with resistance against sorghum shoot fly, can be exploited after sufficient testing under AICRP trials.ThesisItem Open Access A STUDY ON ALLELIC RELATIONSHIP AND IDENTIFICATION OF SNP MARKERS LINKED TO FUSARIUM WILT RESISTANCE GENES IN CASTOR (Ricinus communis L.)(PROFESSOR JAYASHANKAR TELANGANA STATE AGRICULTURAL UNIVERSITY, 2022-01-07) MOHAN RAO, M. D.; SUJATHA, MCastor is an important non-edible oilseed crop having multifarious industrial applications. Its production is hampered by several biotic stresses among which wilt disease caused by Fusarium oxysporum f.sp. ricini is a major one. Being a soil borne disease, management of wilt is difficult by chemical control. Utilization of available wilt resistance sources in developing resistant varieties and hybrids remains the only viable option for management of wilt problem in castor. Though several wilt resistant lines have been identified, the genetics of resistance in those lines are not known. This study was undertaken to decipher the inheritance patterns of wilt resistance genes in a set of resistant lines, understand the allelic relationship among the resistance genes and identify markers linked to wilt resistance genes in different resistance sources. In order to study the inheritance pattern of wilt resistance in a set of nine resistant lines viz., 48-1, AP-48, AP-56, AP-70, AP-111, AP-125, AP-127, AP-156 and AP-163, F1, F2 and BC1F1 populations were generated by crossing the resistant lines with the susceptible line JI-35. The parents, F1s and segregating populations were screened in the wilt sick plot. Screening results indicated that wilt resistance was controlled by single recessive gene in 48-1 and AP-156; while it was controlled by single dominant gene in AP-56, AP-111 and AP-125; two recessive genes with complementary interaction controlled wilt resistance in AP-48, AP-163 and AP-70; and two dominant complementary genes controlled wilt resistance in AP-127. The inheritance patterns of resistance in different sources would help in devising the breeding strategies to incorporate the resistance into breeding lines of castor. The allelism test was carried out to study the allelic relationship between wilt resistance genes from six different resistant lines viz., 48-1, AP-48, AP-56, AP-111, AP 125 and AP-156. The F1s and F2 populations of six resistant × resistant crosses were screened in the wilt sick plot. Allelism test results revealed that the recessive gene in 48-1 and AP-156 were either allelic or tightly linked and were allelic to one of the two recessive genes in AP-48. The dominant genes in AP-56 and AP-111 were either allelic or tightly linked while the dominant gene in AP-125 was non-allelic to the dominant gene in AP-56 and AP-111. As the wilt resistance in castor is predominantly controlled by major genes, the resistance is prone for breakdown. The diverse resistant sources characterized in this study can be utilized in the breeding programmes to diversify the resistance genes so that pathogen evolution may be slowed down. Durable resistance in the cultivars can be achieved by pyramiding multiple resistance genes. In an attempt to validate a set of SNP loci putatively associated with wilt resistance identified through earlier QTL and association mapping studies, the SNPs were converted into KASP assays and subsequently used to genotype six F2 and three BC1F1 populations segregating for wilt resistance. The SNP loci flanking the QTL linked to wilt resistance Rc_28694-84511 (chromosome-7) co-segregated with wilt resistance in the F2 population of 48-1 × JI-35 and predicted the resistance and susceptible phenotypes with 91.7% accuracy. The SNP marker Rc_29706-482910, which is located nearer to Rc_28694-84511, predicted the resistant and susceptible phenotypes of (AP-156 × JI-35) F2 individuals with 92.2% accuracy. The same marker predicted the resistant phenotype in the segregating populations generated using AP-48 and AP-163 with 91.9 to 93.9% accuracy. Thus, the same region was implicated for wilt resistance in four resistant carrying recessive genes for resistance. The SNP marker Rc_29609-103709 on chromosome-8 predicted the resistant and susceptible phenotypes of individuals in segregating population generated using AP-56 and AP-111 (both carrying dominant genes for resistance) with 82.5 to 89.8% accuracy. Thus, this study validated the presence of two major wilt resistance gene clusters on chromosome-7 and chromosome-8, which can be further explored through fine mapping and map-based cloning. The markers linked to resistance genes would help integrating MAS in wilt resistance breeding. Towards development of near isogenic lines (NILs) in the background of the susceptible line JI-35, three backcross populations viz., [(JI-35 × AP-42) × JI-35] BC3F1, [(JI-35 × AP-156) × JI-35] BC2F1 and [(JI-35 × 48-1) × JI-35] BC1F1 were generated during the course of this study. Once developed, the NILs will be useful resources for genetic, molecular and pathological studies on Fusarium wilt in castorThesisItem Open Access A STUDY ON ALLELIC RELATIONSHIP AND IDENTIFICATION OF SNP MARKERS LINKED TO FUSARIUM WILT RESISTANCE GENES IN CASTOR (Ricinus communis L.)(PROFESSOR JAYASHANKAR TELANGANA STATE AGRICULTURAL UNIVERSITY, 2022-01-07) MOHAN RAO, M; SUJATHA, MCastor is an important non-edible oilseed crop having multifarious industrial applications. Its production is hampered by several biotic stresses among which wilt disease caused by Fusarium oxysporum f.sp. ricini is a major one. Being a soil borne disease, management of wilt is difficult by chemical control. Utilization of available wilt resistance sources in developing resistant varieties and hybrids remains the only viable option for management of wilt problem in castor. Though several wilt resistant lines have been identified, the genetics of resistance in those lines are not known. This study was undertaken to decipher the inheritance patterns of wilt resistance genes in a set of resistant lines, understand the allelic relationship among the resistance genes and identify markers linked to wilt resistance genes in different resistance sources. In order to study the inheritance pattern of wilt resistance in a set of nine resistant lines viz., 48-1, AP-48, AP-56, AP-70, AP-111, AP-125, AP-127, AP-156 and AP-163, F1, F2 and BC1F1 populations were generated by crossing the resistant lines with the susceptible line JI-35. The parents, F1s and segregating populations were screened in the wilt sick plot. Screening results indicated that wilt resistance was controlled by single recessive gene in 48-1 and AP-156; while it was controlled by single dominant gene in AP-56, AP-111 and AP-125; two recessive genes with complementary interaction controlled wilt resistance in AP-48, AP-163 and AP-70; and two dominant complementary genes controlled wilt resistance in AP-127. The inheritance patterns of resistance in different sources would help in devising the breeding strategies to incorporate the resistance into breeding lines of castor. The allelism test was carried out to study the allelic relationship between wilt resistance genes from six different resistant lines viz., 48-1, AP-48, AP-56, AP-111, AP 125 and AP-156. The F1s and F2 populations of six resistant × resistant crosses were screened in the wilt sick plot. Allelism test results revealed that the recessive gene in 48-1 and AP-156 were either allelic or tightly linked and were allelic to one of the two recessive genes in AP-48. The dominant genes in AP-56 and AP-111 were either allelic or tightly linked while the dominant gene in AP-125 was non-allelic to the dominant gene in AP-56 and AP-111. As the wilt resistance in castor is predominantly controlled by major genes, the resistance is prone for breakdown. The diverse resistant sources characterized in this study can be utilized in the breeding programmes to diversify the resistance genes so that pathogen evolution may be slowed down. Durable resistance in the cultivars can be achieved by pyramiding multiple resistance genes. In an attempt to validate a set of SNP loci putatively associated with wilt resistance identified through earlier QTL and association mapping studies, the SNPs were converted into KASP assays and subsequently used to genotype six F2 and three BC1F1 populations segregating for wilt resistance. The SNP loci flanking the QTL linked to wilt resistance Rc_28694-84511 (chromosome-7) co-segregated with wilt resistance in the F2 population of 48-1 × JI-35 and predicted the resistance and susceptible phenotypes with 91.7% accuracy. The SNP marker Rc_29706-482910, which is located nearer to Rc_28694-84511, predicted the resistant and susceptible phenotypes of (AP-156 × JI-35) F2 individuals with 92.2% accuracy. The same marker predicted the resistant phenotype in the segregating populations generated using AP-48 and AP-163 with 91.9 to 93.9% accuracy. Thus, the same region was implicated for wilt resistance in four resistant carrying recessive genes for resistance. The SNP marker Rc_29609-103709 on chromosome-8 predicted the resistant and susceptible phenotypes of individuals in segregating population generated using AP-56 and AP-111 (both carrying dominant genes for resistance) with 82.5 to 89.8% accuracy. Thus, this study validated the presence of two major wilt resistance gene clusters on chromosome-7 and chromosome-8, which can be further explored through fine mapping and map-based cloning. The markers linked to resistance genes would help integrating MAS in wilt resistance breeding. Towards development of near isogenic lines (NILs) in the background of the susceptible line JI-35, three backcross populations viz., [(JI-35 × AP-42) × JI-35] BC3F1, [(JI-35 × AP-156) × JI-35] BC2F1 and [(JI-35 × 48-1) × JI-35] BC1F1 were generated during the course of this study. Once developed, the NILs will be useful resources for genetic, molecular and pathological studies on Fusarium wilt in castorThesisItem Open Access TRANSCRIPTOME PROFILING AND ANALYSIS OF Fe AND Zn RESPONSIVE GENES IN PEARL MILLET (Pennisetum glaucum L.)(PROFESSOR JAYASHANKAR TELANGANA STATE AGRICULTURAL UNIVERSITY, 2022-07-07) ANJALI, C; VANISRI, SMicronutrient malnutrition because of Fe and Zn deficiencies is posing a serious health issue in developing countries worldwide. To combat the micronutrient-based hidden hunger, biofortification is an efficient and cost-effective method to enhance the nutrient contents of crops by breeding techniques. Spearheaded by the HarvestPlus Program of Consultative Group for International Agricultural Research (CGIAR), global crop biofortification research was initiated and focused primarily on seven major staple crops including pearl millet, targeting three important micronutrients (Fe, Zn, and vitamin A). Pearl millet (Pennisetum glaucum L.) is a staple for millions of families in dryland tropics and reported varying concentrations of Fe and Zn in germplasm. Studies on the functional characterization of differentially expressed genes (DEGs) and their dynamic role in Fe and Zn pathways will accelerate the biofortified cultivar development. Transcriptome profiling of leaf and root samples of a pearl millet inbred ICMB 1505 under Fe and Zn stress conditions were carried out wherein ten to twelve days old seedlings were exposed to Fe and Zn stress treatments (–Fe–Zn, –Fe+Zn, +Fe–Zn, and +Fe+Zn) for 12 days. Total RNA was extracted from the treated samples followed by cDNA synthesis, cDNA library preparation, and sequencing of the constructed cDNA libraries. A total of 37,093 DEGs under different combinations of leaf and root stress conditions were identified, of which, 7023 and 9996 DEGs were reported in leaf and root stress conditions, respectively. Among the 11,429 unique DEGs, 8605 were annotated to cellular, biological, molecular functions and 458 DEGs were assigned to 39 pathways. The results revealed the expression of major genes related to mugineic acid pathway, phytohormones, chlorophyll biosynthesis, photosynthesis, and carbohydrate metabolism during Fe and Zn starvation in pearl millet. The analysis reported the expression of nicotianamine synthase (NAS), S-adenosyl L- methionine (SAM) synthetase, 2'-deoxymugineic acid (DMA), 2'- deoxymugineic- acid 2'-dioxygenase (IDS3), FER-like transcription factor genes involved in uptake mechanism and oligopeptide transporter 3 (OPT3), Zn-transporter 3 & 5, Zinc induced facilitator 1 (ZIF1), heavy metal ATPases (HMA) genes for transport of Fe and Zn. The study also discussed the expressional changes of several cellular pathway genes and their regulation in Fe and Zn homeostasis under their deficient conditions. The cross-talks between the Fe and Zn provided information on their dual and opposite regulation of key uptake and transporter genes under deficiency conditions. The identified Fe and Zn homeostasis-related genes from the transcriptomic data were annotated onto the pearl millet genome wherein a high number of genes were distributed on chromosome 3 and a low number of genes were distributed on chromosome 7. The orthologues for the top 42 genes were identified in Oryza sativa, Zea mays and Sorghum bicolor along with their annotations and chromosomal positions in respective crops. Moreover, the orthologues of the uncharacterized genes among the top 42 genes, selected based on gene ontology (GO) terms (involved in Fe and Zn homeostasis) reported being major Fe and Zn uptake (NAS, DMAS) and transporter genes (zinc transporter 4 and 9, YSL transporters) in rice, maize, and sorghum. The orthologues identified in rice, maize, and sorghum aids in the genetic biofortification of nutrient contents in these crops. The gene structures of the identified orthologues were represented which helps in understanding the exon-intron positions and evolutionary changes among the species. The knowledge of the position of the above-identified genes in pearl millet, rice, maize, and sorghum crops can be utilized in the genetic biofortification of these crops for Fe and Zn contents by genetic engineering and other breeding programs. Our results assist in developing Fe and Zn-efficient pearl millet varieties through development of genic-SNPs for Fe and Zn responsive genes and their utilization in an ongoing biofortification breeding program to ameliorate malnutrition in the dryland tropics of South Asia and Sub-Saharan Africa.ThesisItem Open Access PHYSICO-CHEMICAL, FUNCTIONAL AND THERMAL CHARACTERIZATION OF DIFFERENTLY TREATED BARNYARD (Echinochloa esculenta), FOXTAIL (Setaria italica) AND LITTLE (Panicum sumatrense) MILLET FLOURS(PROFESSOR JAYASHANKAR TELANGANA STATE AGRICULTURAL UNIVERSITY, 2022-01-02) SHREEJA, K; HYMAVATHI, T.V.Today's health trends shift from reactive to proactive and from treatment to prevention. There is a growing awareness of the importance of health and nutrition and a growing desire to lead an active and healthy lifestyle. This is driving the consumption of millet with increasing awareness of its health benefits. Epidemiological studies have shown that regular consumption of millets reduces the risk of heart disease, minimizes the onset of type II diabetes, improves the digestive system, lowers the risk of cancer, detoxifies the body, and increases immunity including respiratory health and also Parkinson’s disease. Several initiatives are also being undertaken toward enhancing millet cultivation and consumption to reduce health risks. To improve the consumption of millets to a desirable level, in addition to sufficient production, products of commercial importance should be made available in the market. Flour is a basic ingredient for many products across the world. Earlier studies on pretreated flours were not fully investigated. Thus, the aim of the study was to elucidate the differently treated millet flours in terms of physical, functional, thermal, pasting, rheological, nutritional and antioxidant properties. In the present study, three types of minor millets viz., barnyard (BM), foxtail (FM) xxiii and little millet (LM) were selected. Both the whole and dehulled grains were subjected to soaking, germination, fermentation, dry heating, hydrothermal and extrusion treatments after optimization and converted into flours. The resultant flours were examined for physical, functional, pasting, nutritional and antioxidant properties. Storage stability was studied for 90 days. Overall, there was a significant effect of the treatments in all the grains in the different properties studied. Multivariate analysis revealed that the effect was due to treatments than the grains. The mean values of loose bulk density (LBD), tapped bulk density (TBD), color attributes L*, a*, b*, pH, and sedimentation value (SV) were 0.54 (g/ml), 0.68 (g/ml), 18.37 27.88, 3.20, 6.57 and 1.92ml, respectively. LBD of the whole, whole soaked (WS), germinated, fermented and whole hydrothermal (WHT) flours were lower than their respective means. WS, WHT, whole extruded, fermented, and dry heated flours have lower TBD than their respective means. Evaluation of functional properties indicated that water holding capacity (WHC), water absorption capacity (WAC), water solubility index (WSI), swelling power (SP), and swelling index (SI) were significantly improved by extrusion, hydrothermal treatment, germination, and dry heating. The mean WHC, WAC, WSI, SP, SI, foaming capacity (FC) and oil retention capacity (ORC) were 181.44 (%), 6.61%, 3.89, 318.19, 1.41 (g/dl), 13.42 % and 2.14%, respectively. Apart from untreated, whole soaked and whole dried (WDr) flours exhibited higher foaming capacity (FC%) followed by their dehulled counterparts. Foaming was not observed in extruded flours. Whole extruded (WE) and dehulled soaked (DS) flours exhibited highest and lowest ORC, respectively. Measurements of gelatinization temperature revealed a significant effect of treatments and grain on the gelatinization temperature of the millet flours. There was no significant effect of grains on Tc and enthalpy. Hydrothermally treated (HT) followed by, extruded and soaked exhibited the lowest onset, peak and conclusion temperature, while the dehulled fermented flours (DFF) were highest. Enthalpy was lowest in germinated flours (GFFs) and highest in dehulled dry heated flours (DDrF) (P< 0.05). Extrusion significantly improved the mean pasting viscosity (PV) from 244.80 to 4034.33 cP in whole and 227.87 to 3606.33 cP in dehulled flours followed by germinated flours (244.80 to 547.53). DSF, DDrF, DHTF, DFF, and dehulled flours exhibited similar (P<0.05) high peak viscosity (PKV). Extruded followed by germinated flours have very low PKV. Break down viscosity (BV) was significantly high in DDrF, DFF at par with dehulled untreated flours. DSF followed by WHTF and DHTF followed by DFF showed xxiv very high holding viscosity (HV) and Final viscosity (FV) than others, while those of germinated and extruded flours were lower than others. Setback viscosity (SV) of extruded flours, DFF, WHTF and DHTF was significantly higher than others, with no difference among them. Storage and loss modulus were increased in all the treated whole barnyard flour doughs except extruded dough. Among all the millet flours, extruded flours exhibited the lowest G″ and G′. Significantly higher ash, protein, and crude fibre content was in WEF, DHTF and DDrF, respectively. No significant effect of treatments on carbohydrate and fat content. More remarkable improvement was in whole grain treatments than that of dehulled grain. Zinc (6.93 mg/100 g), iron (4.93 mg/100 g), and calcium (42.88 mg/100 g) contents were enhanced in DSF, DSF and Germinated flours, respectively (p<0.05), while extrusion improved potassium levels. Dry heating lowered the protein content. HTF and WFF increased TFC, while that of TPC and tannins increased in extruded flours, which was reflected in lower IC50 and increased DPPH % inhibition values. Dehulled treated flours reduced a higher percentage of oxalates (78.2 to 128.5%) than whole treated (50.6 to 72.2%). Storage of all the flours did not adversely effect the shelf life. Thus, the study demonstrated that treated flours can be developed with enhanced nutrition, physical, functional, thermal, and rheological properties. Further studies on the in vitro digestibility and glycemic index would help promote these flours as functional ingredient.ThesisItem Open Access IDENTIFICATION OF CANDIDATE SNPs FOR BROWN PLANTHOPPER RESISTANCE AND INTROGRESSION INTO ELITE RICE VARIETY TELANGANA SONA(PROFESSOR JAYASHANKAR TELANGANA STATE AGRICULTURAL UNIVERSITY, 2021-01-07) ISHWARYA LAKSHMI, V.G; SREEDHAR, MRice (Oryza sativa L.), one of the world's most important staple food crops is attacked by insect pests of which Brown Planthopper (BPH) is notorious that can cause huge destruction of plants. The present investigation entitled “Identification of candidate SNPs for brown planthopper resistance and introgression into elite rice variety Telangana Sona” was undertaken to study the BPH resistance reaction of genotypes, identifying and designing candidate SNPs related to BPH resistance, validating the germplasm panel with the designed and available SNPs followed by introgression of resistance into the background of Telangana Sona using SSRs and SNPs. A panel comprising of 110 genotypes with five checks were evaluated for BPH resistance using the Standard Seedbox Screening Technique (SSST) in Augmented Randomized Complete Block Design twice during both Kharif 2018 and Rabi 2018-19. The response of genotypes to BPH screening indicated varied levels of resistance reaction that identified 19 genotypes including nine MAGIC lines, six wild species, three gene differentials and a landrace to be resistant with a damage score ranging between 1.0 and 3.0. Single marker analysis detected significant genetic associations (P <0.05) among ten polymorphic markers that were associated with Bph17(t), Bph3, Qbph10, QBph3, Bph33(t) and qBph-3-1 indicating the possibility of these genes/QTLs governing the resistance in the genotypes. Genetic diversity analysis differentiated the genotypes into seven clusters with a clear-cut demarcation of the resistant and susceptible genotypes. Genome-wide Association Studies (GWAS) using GAPIT (Genetic Association and Prediction Integrated Tools) analysis with BLUPs (Best Linear Unbiased Prediction) and cured polymorphic SNPs of 391 MAGIC lines identified 23 significant SNP variants from GLM model. In addition to these, 31 SNPs associated with 13 stress related genes previously detected by Vanisri et al. (2020a) from FarmCPU model were also considered. Of these 54 SNPs, 20 SNPs were selected to cover all 13 annotated genes representing functional defense-related mechanism for which KASP primers were designed. These designed SNPs along with SNPs specific to Bph17, Bph32 and Bph9 genes available with HTPG (High Throughput Genotyping Project) were used for validating the panel of genotypes for BPH resistance. MAGIC lines, M201, M272, M344 and gene differentials RathuHeenati and RathuHeenati accession were found to be having the favorable alleles for both Bph17 and Bph32 genes which could be useful as donors for transferring BPH resistance into popular varieties with marker assisted selection using SNPs. Six designed SNPs, snpOS00912, snpOS00915, snpOS00922, snpOS00923, snpOS00927 and snpOS00929 were found to work efficiently in distinguishing the genotypes into BPH resistant and susceptible clusters which could be further validated in mapping populations to confirm their efficiency as functional markers. Two lines (M229, 10-3) identified as resistant to BPH from SSST, were used for introgressing resistance into Telangana Sona (TS) using the designed SNPs and gene specific SSR markers. Genes/QTLs, Bph33(t) and qBph-3-1 were found to be present in MAGIC line M229, while, Bph33(t) got validated in landrace 10-3 as detected by SSRs. With respect to the available SNPs for Bph17(t) and Bph32, the donor parents did not contain the favorable alleles for these markers, while four of the designed SNPs i.e., snpOS00915, snpOS00920, snpOS00922 and snpOS00923 were polymorphic and could distinguish the parents based on BPH reaction. These designed SNPs were specific to biotype 4 indicating that these four designed SNPs can be inferred as novel loci conferring resistance in the donors as they were slightly away from the reported genes/QTLs on chromosome 1 (Bph38(t), Bph33(t)) and chromosome 6 (Bph32). Hybridization program was initiated to cross the two donor parents with the recurrent parent Telangana Sona to develop F1’s which were confirmed for their hybridity using the foreground SSRs and designed SNPs. BC1F1 populations of both the crosses were developed by backcrossing the F1’s with Telangana Sona. Gene-specific SSR markers and designed SNPs were used for foreground selection which detected two plants (BC1(a)-15, BC1(a)-19) positive for qBph-3-1, nine plants (BC1(a)-2, BC1(a)-4, BC1(a)-7, BC1(a)-9, BC1(a)-14, BC1(a)-16, BC1(a)-18 BC1(a)-19, BC1(a)-20) for Bph33(t) and one plant (BC1(a)-19) positive for both qBph-3-1 and Bph33(t) genes/QTLs from TS//TS/M229, while all the six BC1F1 plants from TS/10-3 were positive for Bph33(t) gene. Eleven backcross plants from TS//TS/M229 and all the six plants from TS//TS/10-3 were phenotypically resistant to BPH as detected from Modified Mass Tiller Screening Method (MMTS). Few BC1F1 plants from TS//TS/M229 had phenotypic resistance in spite of not being positive for qBph-3-1 and Bph33(t) genes/QTLs indicating the presence of novel genes/QTLs governing BPH resistance, for which mapping is desired, while all the BC1F1 plants obtained from TS//TS/10-3 were resistant to BPH both phenotypically and genotypically. The backcross lines were also evaluated for yield and related traits. From the present study, it can be concluded that the backcross lines, BC1(a)-19 and BC1(a)-14 from TS//TS/M229 and BC1(b)-2, BC1(b)-4, BC1(b)-5 from TS//TS/10-3 exhibited superior performance for BPH resistance and yield related traits. Along with resistance, these had early duration, more productive tillers, longer panicles, more number of filled grains, test weight and yield better than the recurrent parent (Telangana Sona) in addition to grain type similar to Telangana Sona and thus, can be used for complete recovery (approx. 90%) of Telangana Sona with desirable level of field resistance to BPH.
