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ThesisItem Open Access CHARACTERIZATION OF VOLATILE ORGANIC COMPOUND (VOCs) IN AROMATIC RICE GENOTYPE USING HS GC-MS/MS(Indira Gandhi Krishi Vishwavidyalaya, Raipur (C.G.), 2020) Raj Kumar; Prasad, Archana S.; Banerjee, Shubha; Parikh, Mangla; Saxena, R. R.The aromatic or fragrant rice sold for higher price has special characteristics. It can be found and sold in the local or export markets at a higher price than conventional rice. Among them, the aromatic cultivars with the highest commercialization globally are the basmati rice of Indian origin and the jasmine rice of Thai origin (Sakthivel et al, 2009). The distinctive odour of aromatic rice is primarily attributed to 2-acetyl-1-pyrroline (2AP), referred to as the popcorn aroma (Buttery et al, 1982), which has an exceptionally low water threshold of about 0.1 ppb. The distinctive odour of aromatic rice is primarily attributed to 2-acetyl-1-pyrroline (2AP), referred to as the popcorn aroma (Buttery et al, 1982), which has an exceptionally low water threshold of about 0.1 ppb. As aroma synthesized in aerial plant parts and then transported to mature grains, it will be insightful to further track the VOCs in different development stages including mature grain. Thus in the present study a total 10 aromatic genotypes of Chhattisgarh state, a national aromatic and a non-aromatic genotype characterized earlier, was selected for characterization of Volatile organic compound (VOCs) using HS/GC-MS/MS and biochemical charectrization under three replication in Kharif 2019-20 seasons (Table 3.1).The diifernt methods used for comparative volatile profiling and biochemical chareterization involved Comparative Volatile Organic compound profiling using advance technology of Head-space configured with GC-MS/MS (Shimadzu Model Tq8040) for automated extraction of volatile organic compounds could be sent for GC/MS analysis properly and Biochemical characterization of Aromatic rice genotypes by processing of samples for estimation of proline content in aromatic rice genotypes of Chhattisgarh with the help of standard protocol of Bates et al, 1973.The identification and profiling of VOCs carried out at two developmental stages Dough and mature grain stage. From the complete profile total 117 compounds are identified out of which 16 are VOCs responsible for pleasant aroma in selected rice genotypes belongs to five Functional Classes as Alcohol ,Aldehyde,Furans,N-Hetrocyclic rings and Terpenes are also reported as aroma compounds which are processed for the further analysis. Hexanal, Silanediol-dimethyl, Heptanal, Octanal, Nonanal, Benzeneacetaldehyde, Decanal, Pyridine-2,4,6 trimethyl and Azulene were identified as major compounds for Aroma in Rice.Silanediol-dimethyl of Alcohol F.G contributed maximum in both developmental stages either aromatic or non-aromatic rice genotypes followed by Aldehyde, N-Hetrocyclic rings, Furans and Terpenes to some extent. The proline content was calculated for local rice cultivars and showed no association with the 2AP content. (Table 4. 2.1 and Figure 4.2.2However, the proline level tends to be correlated with environmental stress in the field of rice cultivation and even storage conditions are also responsible for variations in the quality of rice aroma.All the selected rice genotypes showed proline content in grains ranging (0.34-5.1 µmol/g tissue) after calculating with the help of standard curve. The paddy production has been increased from 500-780 MT in 2009-18. Highest price was for export of Japonica Rice close to it, was Aromatic Rice. Export quantities of of scented rice is less than non-scented rice but export prices of scented rice is comparatively more than non-scented rice, so there is an opportunity of growing more scented rice (www.fao.org). The short grain aromati rice variety of Chattisgarh more or less equivqlent to pioneer basmati rice among all fragrant rice variety which fetch better price to the local farmers of chhatisgarh in the international export market and Such as a distinctive and appealing scent, which are the deciding factors for its commercialization and the consumer's acceptance. On other hand its aroma content majorly 2AP can be used artificially to make different types of aromatic products.ThesisItem Open Access TRANSCRIPTOME CHARACTERIZATION TO IDENTIFY PUTATIVE GENES RELATED TO AMINO ACID BIOSYNTHESIS PATHWAY AND D-AMINO ACID METABOLISM IN LATHYRUS SATIVUS(Indira Gandhi Krishi Vishwavidyalaya, Raipur, 2020) Supriya, Nayak; Verulkar, S.B.Grasspea (Lathyrus sativus L.) is considered as a hardy and wonderful crop owing to its ability to withstand abiotic stress like drought, waterlogging, salinity and cold conditions and some insect herbivory. It can be grown as mixed crop or as “utera” crop with minimal agronomical inputs. Although it is highly nutritious but its cultivation is highly restricted due to its association with a neurotoxic disorder – Neurolathyrism which causes paralysis of lower limbs. The neurolathyrism is caused by a non-protein amino acid β-N-oxalyl-L-α,β-diaminopropionic acid (β-ODAP). β-ODAP is considered to form from the substrate o-acetyl serine (OAS) (Malathi et al.,1970). The immediate precursor of β-ODAP is the BIA which is formed from a ringed structure isoxazolin-5-one. β-ODAP biosynthesis also follows a cyanide detoxification pathway which is supposed to be interlinked with nitrogen and sulphur metabolism pathway. Although the β-ODAP biosynthesis pathway was investigated long back but the genes associated with it are still not known. So in a view to identify the putative genes related to amino acid biosynthesis pathway in Lathyrus sativus , functional annotation of transcripts of Mahateora and RLK-1950 was performed. To functionally annotate the transcriptome data of Mahateora and RLK-1950, a new pipeline MapMan Mercator was approached. This pipeline was specially designed to functionally categorize the orphan plant omics data. The Mercator output (excel sheet) functionally described total 94,793 transcripts and assigned a specific functional category to the 20,562 transcripts in to 34 functional category. This pipeline provided us with more functionally described orphan Lathyrus sativus transcripts as compared to previously approached CLC Genomics Workbench which described functions to only 28,132 transcripts. The Mercator output (excel sheet) was screened was enzymes related to the amino acid biosynthesis, cyanide metabolism, ROS metabolism and d-amino acid metabolism pathways. Among amino acid biosynthesis, alanine aminotransferase, Cysteine synthase, asparagines metabolism and ornithine decycloaminase. Alanine aminotransferase was selected as earlier study suggested a differential correlation between L-alanine synthase and β-ODAP. Cysteine synthase was selected due to its involvement in the cyanide metabolism pathway which leads to formation of Cyanoalanine from OAS which subsequently forms isoxazolin-5-one(precursor for BIA) via asparagine. Consequently, Cyanoalanine synthase, asparaginase and asparagine synthase is also selected. The selection of ROS is dependent due to the fact that whenever the content of ROS increases in the leaves, β-ODAP content reduces significantly and vice-vesa (Jiao et al., 2011). One enzyme associated with d-amino acid namely D-aminoacyl-tRNAdeacylase or D-Tyr-tRNA(Tyr) deacylase was selected due to the fact that β-ODAP is a d-amino acid. Also the cyanoamino pathway from KEGG showed interrelation between Cyanoalanine synthase, D-aminoacyl-tRNAdeacylase and Cysteine synthase. Consequently, 17 transcripts were selected to assess the expression of primers designed from corresponding transcripts in different genotypes of Lathyrus.ThesisItem Open Access STANDARDIZATION OF ALKALINE AND ENZYMATIC TECHNIQUES FOR PURIFICATION OF PROTEIN ISOLATES AND PROCESSED PRODUCTS FROM DIFFERENT VARIETIES OF RICE(Indira Gandhi Krishi Vishwavidyalaya, Raipur, 2020) Kajal Kiran; Verulkar, S.B.Rice is consumed as a staple food in most of the countries of the world. It is mainly a starchy food with less of proteins, lipids, minerals, etc. Consumption of unmilled rice has its own advantages and disadvantages. However, people all over the world prefer milled and well polished rice over un-milled rice or brown rice. Processing of rice grains removes the highly nutritional rice bran layer. The milled rice, otherwise known as white rice, has less of protein compared to the brown rice. Protein isolates are prepared nowadays as protein supplements for the busy and fast moving human life. However, difficulties in the processing, cause the availability of rice protein products less easy. Rice protein in the endosperm is associated tightly on the surface of starch granules creating difficulty in the protein removal. This is also the reason that isolation of rice starch is more costly compared to other starches. Starch is essential for its contribution of calorie to the human diet. Rice varieties differ greatly in their genetic background, granule size, composition and gelatinization which may have an effect on the properties of the isolated starches. Ethanol is an organic compound with the chemical formula C2H5OH. The demand for ethanol has risen in recent years because of its widespread use in chemical, potable and motorfuel industries. Ethanol is produced either by synthetic chemical method or by natural fermentation. A large number of micro-organisms have the capability to convert sugars into ethanol under anaerobic conditions. The waxy nature of rice is preferred in few parts of the world and the waxy gene has been the primary objective of selection during the domestication and subsequent diversification of the rice crop. So, selection of rice varieties using markers can be a work of help for such purposes. Increasing temperature is one of the reasons for reduced rice yield. Heat stress damages the membrane, inactivates enzymes in the chloroplasts and mitochondria, denaturates or degrades proteins, impairs protein and carbohydrate synthesis, productes new proteins, and inhibits carbon metabolism. Thus to meet up the requirement of increasing population, improved varieties of rice with greater heat tolerance are required. The expression analysis of genes responsible for heat stress tolerance may help mitigate this problem. The current study was planned in order to standardize the methods for protein isolation, starch isolation, ethanol production, to study the waxy nature of varieties and to do the expression analysis of heat shock protein genes in rice. Four rice varieties, i.e., Swarna, MTU 1010, RRF – 127 and IB – 91 were considered for protein and starch isolation. Two methods, namely alkaline and enzymatic methods, were followed for protein isolation. The maximum pure protein isolated under alkaline method was highest for RRF - 127 (57.04 %) brown rice under 0.2% NaOH treatment while the lowest amount of pure protein was isolated under 0.3% NaOH treatment from IB - 91 (31.92 %) white rice. The enzymatic method gave the highest percentage of protein isolated from RRF - 127 (61.627 %) and lowest percentage from under the 0.2% NaOH treatment where the lowest extractions were from IB - 91 (40.80 %) white rice under 0.3% NaOH treatment. However, considering both the methods and the two different types of solutions used (0.2% NaOH and 0.3% NaOH), the variety from which the maximum protein could be isolated is RRF – 127 while the variety from which the minimum protein could be isolated is IB – 91. The enzymatic method of protein isolation with 0.2% NaOH treatment is the best method for protein isolation and more protein can be isolated from brown rice as compared to white rice. Highest starch was isolated from Swarna, followed by IB - 91. The least starch was isolated from RRF – 127. Highest starch was isolated from Swarna, followed by IB - 91. The least starch was isolated from RRF – 127. While estimating in the terms of percentage, taking equal amount of brown rice and white rice, white rice has greater starch percentage in comparison to brown rice. This is because white rice has less of protein, lipids, etc than the brown rice. The RM 263 marker is not linked to the waxy gene of the rice as it does not give correlated results with the amylose content of 18 rice varieties taken for the experimentation. Three varieties, namely Swarna, RRF – 127 and MTU 1010 were considered for the expression analysis using heat shock protein genes in rice. The capability of heat stress tolerance is highest in RRF – 127. Swarna has comparatively less heat tolerance. MTU 1010 has the least capability of tolerating heat stress. Upregulation of heat shock protein genes was observed in heat stress conditions as compared to the control conditions in all the three varieties with both OsHSP 26.7 and OsHSP 18 genes. The maximum upregulation was observed in RRF – 127 with OsHSP 26.7. MTU 1010 showed the least fold increase in upregulation with both the genes. Improvements in the standardized methods for protein, starch and ethanol can be made to isolate greater percentages of the respective end products. Marker assisted selection of varieties with waxy nature would make the selection process much easier process than before. Thermo-tolerant rice varieties should be developed to combat the problem of global warming which is increasing day by dayThesisItem Open Access STANDARDIZATION OF ALKALINE AND ENZYMATIC TECHNIQUES FOR PURIFICATION OF PROTEIN ISOLATES AND PROCESSED PRODUCTS FROM DIFFERENT VARIETIES OF RICE(Indira Gandhi Krishi Vishwavidyalaya, Raipur, 2020) Kiran, Kajal; Verulkar, S.B.Rice is consumed as a staple food in most of the countries of the world. It is mainly a starchy food with less of proteins, lipids, minerals, etc. Consumption of unmilled rice has its own advantages and disadvantages. However, people all over the world prefer milled and well polished rice over un-milled rice or brown rice. Processing of rice grains removes the highly nutritional rice bran layer. The milled rice, otherwise known as white rice, has less of protein compared to the brown rice. Protein isolates are prepared nowadays as protein supplements for the busy and fast moving human life. However, difficulties in the processing, cause the availability of rice protein products less easy. Rice protein in the endosperm is associated tightly on the surface of starch granules creating difficulty in the protein removal. This is also the reason that isolation of rice starch is more costly compared to other starches. Starch is essential for its contribution of calorie to the human diet. Rice varieties differ greatly in their genetic background, granule size, composition and gelatinization which may have an effect on the properties of the isolated starches. Ethanol is an organic compound with the chemical formula C2H5OH. The demand for ethanol has risen in recent years because of its widespread use in chemical, potable and motorfuel industries. Ethanol is produced either by synthetic chemical method or by natural fermentation. A large number of micro-organisms have the capability to convert sugars into ethanol under anaerobic conditions. The waxy nature of rice is preferred in few parts of the world and the waxy gene has been the primary objective of selection during the domestication and subsequent diversification of the rice crop. So, selection of rice varieties using markers can be a work of help for such purposes. Increasing temperature is one of the reasons for reduced rice yield. Heat stress damages the membrane, inactivates enzymes in the chloroplasts and mitochondria, denaturates or degrades proteins, impairs protein and carbohydrate synthesis, productes new proteins, and inhibits carbon metabolism. Thus to meet up the requirement of increasing population, improved varieties of rice with greater heat tolerance are required. The expression analysis of genes responsible for heat stress tolerance may help mitigate this problem. The current study was planned in order to standardize the methods for protein isolation, starch isolation, ethanol production, to study the waxy nature of varieties and to do the expression analysis of heat shock protein genes in rice. Four rice varieties, i.e., Swarna, MTU 1010, RRF – 127 and IB – 91 were considered for protein and starch isolation. Two methods, namely alkaline and enzymatic methods, were followed for protein isolation. The maximum pure protein isolated under alkaline method was highest for RRF - 127 (57.04 %) brown rice under 0.2% NaOH treatment while the lowest amount of pure protein was isolated under 0.3% NaOH treatment from IB - 91 (31.92 %) white rice. The enzymatic method gave the highest percentage of protein isolated from RRF - 127 (61.627 %) and lowest percentage from under the 0.2% NaOH treatment where the lowest extractions were from IB - 91 (40.80 %) white rice under 0.3% NaOH treatment. However, considering both the methods and the two different types of solutions used (0.2% NaOH and 0.3% NaOH), the variety from which the maximum protein could be isolated is RRF – 127 while the variety from which the minimum protein could be isolated is IB – 91. The enzymatic method of protein isolation with 0.2% NaOH treatment is the best method for protein isolation and more protein can be isolated from brown rice as compared to white rice. Highest starch was isolated from Swarna, followed by IB - 91. The least starch was isolated from RRF – 127. Highest starch was isolated from Swarna, followed by IB - 91. The least starch was isolated from RRF – 127. While estimating in the terms of percentage, taking equal amount of brown rice and white rice, white rice has greater starch percentage in comparison to brown rice. This is because white rice has less of protein, lipids, etc than the brown rice. The RM 263 marker is not linked to the waxy gene of the rice as it does not give correlated results with the amylose content of 18 rice varieties taken for the experimentation. Three varieties, namely Swarna, RRF – 127 and MTU 1010 were considered for the expression analysis using heat shock protein genes in rice. The capability of heat stress tolerance is highest in RRF – 127. Swarna has comparatively less heat tolerance. MTU 1010 has the least capability of tolerating heat stress. Upregulation of heat shock protein genes was observed in heat stress conditions as compared to the control conditions in all the three varieties with both OsHSP 26.7 and OsHSP 18 genes. The maximum upregulation was observed in RRF – 127 with OsHSP 26.7. MTU 1010 showed the least fold increase in upregulation with both the genes. Improvements in the standardized methods for protein, starch and ethanol can be made to isolate greater percentages of the respective end products. Marker assisted selection of varieties with waxy nature would make the selection process much easier process than before. Thermo-tolerant rice varieties should be developed to combat the problem of global warming which is increasing day by day.ThesisItem Open Access Identification and Validation of Gene Tagged SNP Markers linked to Yield Related Traits in Rice.(Indira Gandhi Krishi Vishwavidyalaya, Raipur (C.G.), 2020) Verma, Rupesh Kumar; Prasad, Archana S.Rice is the most popular agricultural crop and is the main hub of many developing countries of Asia on the continent. It also used as a food source in the continent of Africa Rice production should be increased to meet the growing need of rice to feed the growing population increased genetic efficiency is an important measure for the efficiency of rice production. It is important to identify the genes to be improved to produce fresh mega rice through breeding programs based on rapid molecular marker for crop development. In this study, F3 population lines were evaluated during Kharif season 2019 -2020 under irrigated condition and phenological data for 13 yield related traits were recorded. On the basis of mean performance, significant variation was observed in all the traits under irrigated condition. SNP markers flanking 2 yield related traits reported to explain high percentage phenotypic variation for their respective traits in 7 population from a cross MTU1010×WR151 and Swarna×RRE78, Swarna×WR1, Swarna×RRF78, MTU1010×WR162, MTU1010×WR-116, MTU1010×DJOGLON were used for validation in F3 population single marker analysis was carried out by single factor ANOVA. A total of their amplification band were obtained indicated the presence of two alleles for each parent. For DEP-1 (T/A) (Middle exon/ 5'- exon) the a band of size 600bp indicated expression in turn amplification of DEP-1 gene, where as a band of size 520bp indicated presence T/A SNP allele and 380bp showed SNP Allele. Similarly, for GHD-7 (G/A) (First exon) gene band of size 580bp indicated amplification of targeted gene, and size of 550bp and 400bp indicated expression and amplification of SNP alleles. In the F3 population set, the marker DEP-1 showed a strong association of SWARNA× RRF78 with grain dimension traits such as flag leaf breath, biological yield, grain yield, explaining 0.6%, 23.12 %, 23.13 %, and MTU1010×WR-162 grain dimension 50% flowering, plant height, flage leaf length, grain length, grain breath, grain weight 100 seed, explaining 6.99%,6.49%,7%, 9.01% 5.9%, 4.51% of phenotypic variance, respectively. The marker-trait association for DEP-1 and were also found to be significant. Similarly In F3 population, the GHD-7 gene based marker MTU1010×WR-151 was found to be strongly associated with 50% flowering, no of tillers, grain length, grain weight 100 seed, explaining 0.17%, 14.05%, 0.34%, 2.2%, and SWARNA×RRF78 grain dimension 50% flowering, no of tillers, explaining 0.9%, 2.59%, and MTU1010×WR-116 grain dimension grain weight, explaining 0.3% of the phenotypic variance respectively at significant P values.ThesisItem Open Access Identification and Validation of Gene Tagged SNP Markers linked to Yield Related Traits in Rice(Indira Gandhi Krishi Vishwavidyalaya, Raipur, 2020) Verma, Rupesh Kumar; Prasad, Archana S.; Chandel, Grish; Dongre, Prabha Rani; Saxena, R.R.; Verulkar, S.B.Rice is the most popular agricultural crop and is the main hub of many developing countries of Asia on the continent. It also used as a food source in the continent of Africa Rice production should be increased to meet the growing need of rice to feed the growing population increased genetic efficiency is an important measure for the efficiency of rice production. It is important to identify the genes to be improved to produce fresh mega rice through breeding programs based on rapid molecular marker for crop development. In this study, F3 population lines were evaluated duringKharif season 2019 -2020 under irrigated condition and phenological data for 13 yield related traits were recorded. On the basis of mean performance, significant variation was observed in all the traits under irrigated condition. SNP markers flanking 2 yield related traits reported to explain high percentage phenotypic variation for their respective traits in 7 population from a cross MTU1010×WR151 and Swarna×RRE78, Swarna×WR1, Swarna×RRF78, MTU1010×WR162, MTU1010×WR-116, MTU1010×DJOGLON were used for validation in F3 population single marker analysis was carried out by single factor ANOVA. A total of their amplification band were obtained indicated the presence of two alleles for each parent. For DEP-1 (T/A) (Middle exon/ 5'- exon) the a band of size 600bp indicated expression in turn amplification of DEP-1 gene, where as a band of size 520bp indicated presence T/A SNP allele and 380bp showed SNP Allele. Similarly, for GHD-7 (G/A) (First exon) gene band of size 580bp indicated amplification of targeted gene, and size of 550bp and 400bp indicated expression and amplification of SNP alleles. In the F3 population set, the marker DEP-1 showed a strong association of SWARNA×RRF78 with grain dimension traits such as flag leaf breath, biological yield, grain yield, explaining 0.6%, 23.12 %, 23.13 %, and MTU1010×WR-162 grain dimension 50% flowering, plant height, flage leaf length, grain length, grain breath, grain weight 100 seed, explaining 6.99%,6.49%,7%, 9.01% 5.9%, 4.51% of phenotypic variance, respectively. The marker-trait association for DEP-1 and were also found to be significant. Similarly In F3 population, the GHD-7 gene based marker MTU1010×WR-151 was found to be strongly associated with 50% flowering, no of tillers, grain length, grain weight 100 seed, explaining 0.17%, 14.05%, 0.34%, 2.2%, and SWARNA×RRF78 grain dimension 50% flowering, no of tillers, explaining 0.9%, 2.59%, and MTU1010×WR-116 grain dimension grain weight, explaining 0.3% of the phenotypic variance respectively at significant P values.ThesisItem Open Access DOUBLE HAPLOID PRODUCTION AND IN-VITRO SCREENING OF DROUGHT TOLERANCE IN RICE(Indira Gandhi Krishi Vishwavidyalaya, Raipur, 2020) Archana; Jha, Zenu; Verma, Sunil; Bhandarkar, S.; Saxena, R.R.In anther culture γ radiation is the mutation breeding tool used to obtained homozygous lines. The use of γ radiation increases the callus initiation and plant regeneration frequency. The present work was done to study the effect of γ radiation on callus initiation and plant regeneration of Dubraj. The callus was subjected to different doses of γ radiation viz 10,20,40,60,80 Gy along with control (0) respectively. When the radiation level increased, the maximum green callus percent was obtained at 10 Gy (25.6) as compared to control condition (12.7) and further decrease at 40,60 and 80 Gy. Anther culture derived plants, were either haploid homozygous and diploid heterozygous to distinguish between them ploidy analysis was done to establish homozygosity. A flow cytometry method was used for ploidy determination which is an expensive and costly. To identify haploids and diploids in anther culture, cytological study was done in rice with the help of a light microscope which is a cheapest method. Thirty days old callus from haploid and diploid callus were taken for cell and nucleus size measurement. A result shows that nucleus size of haploid cell is nearly half of the diploid cell. Drought decreases total biomass production by causing stress at vegetative and reproductive stagesin rice cultivars. Ricedouble haploid cross Swarna sub 1x IR-159 with a forty-one plant genotypes were screened in vitro at various concentrations of Polyethylene glycol (PEG) 6000-3.0, -5.0, -7.5, -10.0, -12.5 and control (0) bars of water potentialData were recorded on root length and shoot length of 12-15 days old seedling. Among all the double haploid lines, seven lines were found to be drought tolerant. At highest concentration -12.5, the healthy growth was observed with the mean root length (4.3) and shoot length (3.4).The genomic analysis reveals that obtained double haploid lines were genotypically drought tolerant by using SSR markers.ThesisItem Open Access Validation of Gene Tagged Markers of Yield Related Traits In Rice(Indira Gandhi Krishi Vishwavidyalaya, Raipur, 2020) Verma, Sunil Kumar; Prasad, Archana; Saxena, R.R.; Banerjee, Shubha; Sharma, Bhawna; Saxena, R.R.Rice is the most popular agricultural crop and is the main hub of many developing countries of Asia on the continent. It also used as a food source in the continent of Africa Rice production should be increased to meet the growing need of rice to feed the growing population increased genetic efficiency is an important measure for the efficiency of rice production. It is important to identify the genes to be improved to produce fresh mega rice through breeding programs based on rapid molecular marker for crop development. In this study, F2 population lines were evaluated during kharif season 2018 and 2019 under irrigated condition and phenological data for 13 yield related traits were recorded. On the basis of mean performance, significant variation was observed in all the traits under irrigated condition. SNP markers flanking 2 yield related traits reported to explain high percentage phenotypic variation for their respective traits in 4 population from a cross Swarna x RRF-78, MTU1010 X WR-151, MTU1010 X WR-162 and MTU1010 X WR-116 were used for validation in F2 population single marker analysis wascarryed out by single factor ANOVA. A total of their amplification band were obtained indicated the presence of two alleles for each parent. For DEP-1 (T/A) (Middle exon/ 5'- exon) the a band of size 600 bp indicated expression in turn amplification of DEP-1 gene, where as a band of size 520 bp indicated presence T/A SNP allele and 380 bp showed SNP Allele. Similarly, for GHD-7 (G/A) (First exon) gene band of size 580 bp indicated amplification of targeted gene, and size of 550 bp and 400 bp indicated expression and amplification of SNP alleles. In the F2 population set, the marker DEP-1 showed a strong association of SWARNA × RRF-78 with grain dimension traits such as flag leaf breath, biological yield, grain yield, explaining 0.6%, 23.12 %, 23.13 %, and MTU1010 X WR-162 grain dimension 50% flowering, plant height, flage leaf length, grain length, grain breath, grain weight 100 seed, explaining 6.99%,6.49%,7%, 9.01% 5.9%, 4.51% of phenotypic variance, respectively. The marker-trait association for DEP-1 and were also found to be significant. Similarly In F2 population, the GHD-7 gene based marker MTU1010 X WR-151 was found to be strongly associated with 50% flowering, no of tillers, grain length, grain weight 100 seed, explaining 0.17%, 14.05%, 0.34%, 2.2%, and SWARNA X RRF-78 grain dimension 50% flowering, no of tillers, explaining 0.9%, 2.59%, and MTU1010 X WR-116 grain dimension grain weight, explaining 0.3% of the phenotypic variance respectively at significant P values.ThesisItem Open Access Validation of Gene Tagged Markers of Yield Related Traits In Rice(Indira Gandhi Krishi Vishwavidyalaya, Raipur, 2020) Verma, Sunil Kumar; Prasad, Archana; Banerjee, Shubha; Sharma, Bhawana; Saxena, R.R.Rice is the most popular agricultural crop and is the main hub of many developing countries of Asia on the continent. It also used as a food source in the continent of Africa Rice production should be increased to meet the growing need of rice to feed the growing population increased genetic efficiency is an important measure for the efficiency of rice production. It is important to identify the genes to be improved to produce fresh mega rice through breeding programs based on rapid molecular marker for crop development. In this study, F2 population lines were evaluated during kharif season 2018 and 2019 under irrigated condition and phenological data for 13 yield related traits were recorded. On the basis of mean performance, significant variation was observed in all the traits under irrigated condition. SNP markers flanking 2 yield related traits reported to explain high percentage phenotypic variation for their respective traits in 4 population from a cross Swarna x RRF-78, MTU1010 X WR-151, MTU1010 X WR-162 and MTU1010 X WR-116 were used for validation in F2 population single marker analysis wascarryed out by single factor ANOVA. A total of their amplification band were obtained indicated the presence of two alleles for each parent. For DEP-1 (T/A) (Middle exon/ 5'- exon) the a band of size 600 bp indicated expression in turn amplification of DEP-1 gene, where as a band of size 520 bp indicated presence T/A SNP allele and 380 bp showed SNP Allele. Similarly, for GHD-7 (G/A) (First exon) gene band of size 580 bp indicated amplification of targeted gene, and size of 550 bp and 400 bp indicated expression and amplification of SNP alleles. In the F2 population set, the marker DEP-1 showed a strong association of SWARNA × RRF-78 with grain dimension traits such as flag leaf breath, biological yield, grain yield, explaining 0.6%, 23.12 %, 23.13 %, and MTU1010 X WR-162 grain dimension 50% flowering, plant height, flage leaf length, grain length, grain breath, grain weight 100 seed, explaining 6.99%,6.49%,7%, 9.01% 5.9%, 4.51% of phenotypic variance, respectively. The marker-trait association for DEP-1 and were also found to be significant. Similarly In F2 population, the GHD-7 gene based marker MTU1010 X WR-151 was found to be strongly associated with 50% flowering, no of tillers, grain length, grain weight 100 seed, explaining 0.17%, 14.05%, 0.34%, 2.2%, and SWARNA X RRF-78 grain dimension 50% flowering, no of tillers, explaining 0.9%, 2.59%, and MTU1010 X WR-116 grain dimension grain weight, explaining 0.3% of the phenotypic variance respectively at significant P values.
