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Open Access
Genome wide association mapping for stem water soluble carbohydrates in bread wheat (Triticum aestivum L.) under terminal water stress
(CCSHAU, Hisar, 2020-11-26) Gaur, Arpit; Jindal, Yogesh
Genomic assisted breeding is one way to improve drought stress tolerance in wheat. To uncover the genomic regions for grain yield, its attributing traits, water soluble carbohydrates (WSCs) and its components mainly fructans, a comprehensive multidisciplinary study was conducted with a diverse set of 302 bread wheat genotypes. All the genotypes were evaluated under complete irrigation, rainfed and complete water stressed condition for two consecutive crop seasons at three locations (Hisar, Karnal and Baramati). The study indicated significant effect of different water regimes and locations on the expression of traits and as much as 80% of yield reduction was reported under DT. Seven promising drought tolerant verities were identified on the basis of drought susceptibility index. In this study, correlation between yield and studied traits largely varied with environmental and water conditions. However, shorter plant height, longer grain filling duration, days to heading, thousand kernel weight (35-40 g) is the most favorable trait combination for obtaining high grain yield under studied conditions. Positive correlation of WSCs with grain yield was reported only for drought tolerant genotypes. High values of all the genetic components were reported for all the traits within environments, however in pooled over management these values ranged between low to moderate for most of the studied traits due to genotype x environment. Traits like plant height, peduncle length, spikelet per spike and thousand kernel weight were least influenced with genotype x environment interaction thus found suitable for selection of grain yield. Substantial genetic diversity was reported in the studied material which was attributed by each of the studied trait equally. With 35K Axiom array and applying compressed mixed linear model, 431 stable SNPs were identified for 20 studied traits in 1288986.92 cM genomic region of bread wheat. These SNPs explained ca. 30-53% of phenotypic variations. Of 431 stable SNPs 77 were showing pleiotropy for various traits. Five SNPs (AX-94583229, AX-94879209, AX-94759517, AX-94887053 and AX-94964616) for fructans and total non-structural carbohydrates, three (AX-94508292, AX-95257620 and AX-94456473) for days to heading and one (AX-94412521) for plant height were most significant. In functional analysis of significant SNPs, 249 genes were identified which were mainly involved in carbohydrate metabolism and repair mechanism. Only few were reported with direct involvement in stress responses. In an in silco study 104 genes (TaFruc) transcribing into stable protein associated with fructans metabolism were identified. All of these were involved in either hydrolase or transferase activities of glycosyl groups of soluble sugars. These genes were found expressive under different stress conditions, mainly water stress. To facilitate the future breeding programs a total of 189 SSR molecular markers and 60 micro RNAs have been predicted successfully for TaFruc genes.
Open Access
Interaction of Sesbania rhizobia with different species of Sesbania and kharif legumes
(CCSHAU, Hisar, 2020-11) Jeniffer Christeena Masih; Gera, Rajesh
Sesbania is an important wild legume as it can grow in wide range of soils like alkaline, waterlogged and acidic soils. It has high nitrogen fixation properties due to its association with rhizobia in both root and stem nodules. Rhizobia from wild legume like Sesbania may function as excellent plant growth promoting bacteria.Therefore, the present research was carried out to study the interaction of Sesbania rhizobia with different species of Sesbania and kharif legumes. A total of 25 Sesbania rhizobial isolates, which includes five isolates each from Sesbania aculeata, S. sesban, S. grandiflora, S. rostrata (root nodulating and stem nodulating), were included in the present investigation. Out of which 21 rhizobial isolates were obtained from departmental culture collection and 4 rhizobial isolates were isolated from soils collected from different locations of India using trap plant method. All the rhizobial isolates were able to produce IAA and ammonia, however, 92, 48 and 48 % rhizobia had the ability for phosphate solubilization, bacteriocin and siderophore production, whereas, 60% of rhizobia were able to utilize ACC. All the rhizobial isolates showed the presence of nifH and nodC genes. Five rhizobial isolates namely SSKr(ii), SGMg, SAUd(i), SRKr(iv)/r and SRTn/s from each Sesbania species were selected on the basis of different plant growth promoting traits, nodulation efficiency and amplification of nodC and nifH gene, to study their effect on different Sesbania species, mungbean and pigeonpea under sterilized conditions. The rhizobial isolates; SRKr(iv)/r (root nodulation) and SRTn/s (stem nodulation) were found to be most efficient isolates on the basis of nodule number and fresh nodule weight in cross nodulation within Sesbania species and other legumes. These isolates were also tagged with gfp gene to study their colonization on different parts of Sesbania rostrata. Recovery of GFP marked strains from root, root nodules, stem and stem nodules was 95 to100%. However, recovery of gfp marked strains from the surface of root and root nodules varied from 92 to100% while on the surface of stem and stem nodules, it ranged between 25 to 33%. Rhizobial isolate; SRKr(iv)/r showed good nodulation efficiency in all four Sesbania species and pigeonpea as compared to other rhizobial isolates under unsterilized conditions. Nodule occupancy of GFP marked strains; SRKr(iv)/rGFP+ and SRTn/sGFP+ under unsterilized condition was found to be 33-54 and 92% in case of root and stem nodules, respectively of Sesbania rostrata.
Open Access
Biochemical changes in genetic and induced systemic resistance due to infection of Albugo candida (Pers. ex. Lev) in Indian mustard
(CCSHAU, Hisar, 2020-03) Annu; Rathi, A. S
White rust disease incited by Albugo candida is an important disease of Indian mustard (Brassica juncea) in India particularly in northern and North eastern states. Keeping in view the enormous losses caused by the pathogen, the present investigation has been devised on biochemical changes due to infection of Albugo candida in different Indian mustard genotypes, induction of systemic resistance by non conventional chemicals, identification of slow white rusting genotypes and role of micronutrients in defence against white rust under laboratory, screen house and field conditions at Chaudhary Charan Singh Haryana Agricultural University, Hisar. The results of biochemical analysis revealed that total phenolic content, PO, PPO, PAL and chitinase activity were found higher in healthy leaves of resistant genotypes (BIOYSR, EC399299) as compared to moderately resistant genotypes (EC399301, DRMRIJ-31) and susceptible genotypes (RH 30, RH 8812) at 35-40 DAS. There was steep increase in all the biochemical constituents at the time of disease initiation ie. at 45-50 DAS. However, increase was more in inoculated environment as compared to uninoculated environment. At maximum disease severity i.e.70-75 DAS, there was decline in all the biochemical constituents in all genotypes under both the environments. Decrease was more in susceptible genotypes indicating the exhaustion of these biochemical constituents which otherwise consider necessary to trigger genetic resistance through pathways. Efficacy of six non conventional chemicals were tested with different concentrations for induction of resistance against white rust. None of the non conventional chemical completely controlled the white rust disease even at higher concentration of 100 μg/ml. However, a maximum control was also recorded by BTH at 100 μg/ml concentration after six weeks of challenge inoculation by the pathogen followed by SA in both the variety (RH 30 and RH 0749) under screen house conditions. BTH has shown maximum induction of phenolic content, PO, PPO, PAL and chitinase activity at three days after inoculation (DAI) in both the varieties at 100 μg/ml concentration followed by SA. However, MgSO4 has shown minimum induction of these activities at three DAI in both the varieties.Out of 22 Indian mustard genotypes screened for identification of slow white rusting component, 9 genotypes exhibited longer incubation and latent periods, minimum number of pustules per leaf, minimum size of pustule, minimum sporulation, minimum disease severity and minimum AUDPC indicating slow white rusting behaviour. Among the different treatments of mcronutrients alone and in combinations, soil application of Gypsum + Borax + CuSO4 + Na2MoO4 at 30 DAS as basal dose recorded minimum white rust severity of 31.1% and staghead incidence of 4.44% and highest seed yield of 2228.55 kg ha-1 as compared to the 45.83% white rust severity and 6.09% staghead incidence and seed yield of 1779.79 kg ha-1 in control. Maximum number of primary branches per plant (5.83), secondary branches per plant (12.33), main shoot length (90.93 cm), siliquae on main shoot (62.60) and seeds per siliquae (12.53) were also observed with above treatment. Plant height, siliqua length, oil content and 1000 seed weight did not vary significantly among all the treatments.
Open Access
Molecular approaches for detection and forecasting of wheat yellow rust
(CCSHAU, Hisar, 2020-03) Rizwana Rehsawla; Yadav, Neelam R.
Yellow or stripe rust, caused by Puccinia striiformis f. sp. tritici (Pst), is a devastating airborne disease that affects bread wheat in the major wheat growing regions of India. The understanding of the origin, evolution, pathogenicity, avirulence/virulence behaviour of Pst is very important for the development of more effective management strategies to combat the disease spread. To understand inter and intraspecific phylogenetic relationship among Indian Pst pathotypes, multigene sequence analysis was done. Molecular marker study along with sequencing technology was used to collect information, which was more effective than virulence characterization. The molecular diversity analysis among 13 different Pst pathotypes showed two major cluster formations at similarity coefficient of 0.78. Sometimes, all three wheat rust or two rusts in combination occur simultaneously in the field. Under such conditions identification and differentiation of the yellow rust is needed for precise identification and high throughput DNA based detection protocols. To address this problem PCR based markers were developed which can specifically detect and differentiate Pst from two other rust species of Puccinia and other wheat pathogens. DNA-based methods such as conventional PCR have revolutionized plant disease detection; they are not very reliable at asymptomatic stage. Therefore, a simple and reproducible LAMP assay was developed which could detect the pathogen at an early stage i.e. 3rd day of post infection without any visible sign of pathogen attack on the leaf sample using LAMP primers available in public domain. Four novel sets of LAMP primers from ketopantotate reductase gene were also designed for Pst detection which worked successfully. Biosensing by electrochemical and SPR for the detection of yellow rust was also undertaken. Electrochemical based sensing was done using different sequences of Pst specific genes as probes. Linear response over wide DNA concentration range from 10 pg/μl to 115ng/μl was obtained with a high sensitivity, accuracy and reproducibility. The lowest detection limit was observed for microRNA like RNA 1 gene i.e. 10 pg/μl. Electrochemical DNA based biosensing was developed to distinguish between the yellow rust susceptible and resistant wheat genotypes using TaATG8j gene sequence. Linear response over wide DNA concentration range from 1 ng/μl to 50 ng/μl was obtained with detection limit of 4 pg/μl. The SPR biosensor demonstrated high specificity and long shelf life thus promising its application in Pst diagnosis. The developed biosensor exhibited a high sensitivity (0.18°/ (ng/μl)), good linearity, low detection limit (1 ng/μl) and high specificity over a wide concentration range of DNA (1–150 ng/μl).
Open Access
Processing of pumpkin seeds and their utilization in product development
(CCSHAU, Hisar, 2020-06) Neeta Kumari; Sindhu, Sangeeta C.
The present investigation was conducted to evaluate the functional properties and nutrient composition of processed pumpkin seed powder. Processed pumpkin seed powder was used for the development of various value added food products like whole wheat flour biscuit, cookies, Laddoo, Lapsi, bread, buns, Dhokla and eggless muffins. Pumpkin seeds were given four types of treatments i.e. germination, boiling, roasting and fermentation. Processed pumpkin seeds included raw, boiled, roasted, germinated, natural fermented, lactobacillus acidophilus fermentation and lactobacillus rhamnosus fermentation. All processing methods brought significant changes in functional properties of pumpkin seeds. The functional properties of each treated sample differed significantly from each other. Germinated seed powder had highest oil absorption capacity (89.33%), water absorption capacity (276.06%) and swelling power was significantly (P≤0.05) higher in roasted pumpkin seed powder (3.23%). Ash content was highest in roasted pumpkin seed powder whereas crude fat, crude fiber and crude protein were highest in germinated pumpkin seed powder. All the processed sample were significantly (P≤0.05) different from each other. Total minerals (magnesium 11.73, zinc 17.73, iron 22.77 and phosphorus 68.22 mg/100g) were significantly (P≤0.05) higher in germinated pumpkin seed powder as compared to control counterpart. Calcium 164.30 and potassium 1315 mg/100g were significantly (P≤0.05) higher in roasted pumpkin seed powder than any other processed sample. Per cent HCl-extractability of minerals like calcium, magnesium, potassium were significantly (P≤0.05) higher in roasted pumpkin seed powder followed by germinated pumpkin seed powder while iron and zinc were significantly higher in germinated pumpkin seed powder. All the processing methods brought significant (P≤0.05) decrease in phytic acid and polyphenol content. Total plate count of all the processed samples increased with storage. Products like whole wheat flour biscuit, cookies, Laddoo, Lapsi and eggless muffins were developed by replacing (10%, 20% and 30%) of germinated pumpkin seed powder. Dhokla, bread and buns were developed by incorporating various levels (10%, 20% and 30%) of fermented pumpkin seed powder. Total 32 products were prepared and subjected to organoleptic acceptability. The acceptability level of whole wheat flour biscuits, cookies, Laddoo, eggless muffins, Dhokla and Lapsi was up to 30% whereas the acceptability level in case of bread and buns was up to 20%. Products were evaluated for sensory and nutritional parameter. All products depicted good nutritional profile with significant improvement in all the nutrients. Eggless muffins, bread and buns could be stored for 2 days at room temperature 42◦C without any adverse effect on the organoleptic characteristics. Whole wheat flour biscuits and cookies, Laddoo were could be stored for seventy five days safely without adverse effect on organoleptic acceptability. The values for peroxide value and total plate count of all stored products showed an increasing trend with the increased in storage periods. There was a significant (P≤0.05) increase in the knowledge level of trainees after the training at 1% level of significance.
Open Access
Structure-based virtual high-throughput screening for ligands of ecdysone receptor
(CCSHAU, Hisar, 2020) Gagan Rani; Sudhir Kumar
Insects occupy more than 70% of entire kingdom Animalia and are the most successful group o among the organisms dwelling on earth, a set of injurious insects referred as pests. Pests/insects can harm agricultural plants, devour and/or harm harvested crops or transmit diseases to people and animals. These insect pests have several enzymes. hormones receptors, these hormone receptor binds with specific hormone and thus regulates development and also plays a key role in their life processes. If the normal functioning of these enzymes and the receptors are disturbed, their normal life cycle may also be affected thereby causing death of the insect. In the present study we focused only on ecdysone receptor, which is a type of nuclear receptor steroids hormone. The ecdysone receptor (EcR) is a nuclear transcription factor depends upon ligands found in arthropods. The receptor binds to ecdysteroids hormones, which play a major role in reproduction and regulate vital processes such as growth, molting and metamorphism. For activation of gene transcription, a non-covalent heterodimerization of EcR and USP (ultra-spiracle protein) is necessary, that occurs after binding of an agonist to ecdysone receptor. Because of the importance of this hormone receptor in growth, reproduction and metamorphosis of insects, it was considered to be excellent targets for pest control agents and chemical insecticides. In the presents study virtual high-throughput of two libraries of 705,632 (on the basis of substructure) and 4591,276 (clean leads) were carried out for ecdysone receptor. At the same time vHTS of 21 known inhibitors for ecdysone receptor also performed. vHTS results showed that 1,85,785 and 5,15,112 leads had binding energy within the range or lesser than the binding energy of known inhibitors (from both libraries respectively). These selected leads were then subjected to ADME-Tox study, from this 264 and 509 leads were predicted to be non-toxic.
Open Access
Sustainable integrated farming system in Haryana: Problems and prospect
(CCSHAU, Hisar, 2020-03) Ekta; Malik, Joginder Singh
The study was conducted in Yamunanagar and Sirsa districts of Haryana state during the year 2018-19. These districts were purposively selected. Further, four villages from each district were selected, randomly. Further, 22 farmers were selected, randomly from each selected village. Thus, a total number of 176 respondents were interviewed for the present study. The information about respondents‟ socio-personnel traits, need and interest of farmers in adoption of IFS, feasible components of IFS in Haryana, farmer‟s perception regarding the impact of IFS in sustainable agriculture, problems and prospect of small and marginal farmers in different farming systems in Haryana were collected with the help of structured interview schedule. The study indicated that there were 63.07 per cent of respondents who had medium need and interest towards IFS. 31.82 per cent respondent‟s perceived high need and interest towards IFS. Whereas, only 5.11 per cent of the respondents perceived less need and interest towards IFS. In case of IFS components adopted by the respondents, all of the respondents had adopted field crop followed by dairy farming, forestry, horticulture and vegetables etc. Results pertaining to the perception of farmers regarding the impact of IFS in sustainable agriculture found that majority of respondent 51.70 per cent had medium level of perception towards IFS. The study conducted on constraints encountered by the respondents in adoption of IFS revealed that there were 47.73 % of respondents who believed to have medium problems regarding IFS such as Economical, technical, production, marketing and organizational problems. The overall results of IFS prospect perceived by the respondents showed that about 55.11 per cent of the respondents perceived medium to high level of IFS, hence the future of IFS is bright under these circumstances. The study further revealed that most of the respondents believed to have medium level of technical prospect (64.77%), economical prospect (66.48%), social prospect (52.27%) and ecological prospect (53.98%) regarding IFS. They also believed to have low level of entrepreneurial prospect (57.95%). IFS play an important role in reducing of the use of chemical fertilizers by improving the soil health naturally by increasing the soil nutrients, organic carbon and micro-organism in the soil.
Open Access
Quantitative and molecular characterization for heat tolerance in wheat (Triticum aestivum L. em. Thell).
(CCSHAU, Hisar, 2020-02) Anil Kumar; Dhanda, S.S
The present study was carried out to evaluate 96 advanced recombinant inbred lines derived from a cross WH 730 (heat tolerant) and WH 147 (higher yielder, heat susceptible) to find out physio-morphological traits and quantitative trait loci (QTLs) associated with heat tolerance in bread wheat. The experiment was conducted in augmented design and randomized complete block design (RBD) during Rabi season of 2014- 15, 2015-16 and 2016-17 in the Department of Genetics and Plant Breeding, CCS Haryana Agricultural University, Hisar. Analysis of variance revealed significant differences for all traits studied among the RILs. Medium to high values of GCV, PCV, heritability and genetic advance as percent of mean for grain yield per plant, number of grains per spike,100-grain weight, biological yield, canopy temperature depression and cell membrane thermostability under either one or both environments over the years indicating a high scope of selection for these traits for heat tolerance. Grain yield was significantly and positively correlated with number of tillers per plant, harvest index, biological yield per plant, 100-grain weight, canopy temperature depression, cell membrane thermostability and osmotic potential, whereas negative and significantly correlated with days to heading and days to maturity under timely and late sown conditions over the years. Biological yield and harvest index had high direct and indirect effects on grain yield per plant over environment and years, indicating the true relationships with grain yield, in this set of material. Nine promising RILs namely RIL No.46, 22, 40, 43, 57, 19, 48, 81, 71 and 88, out of 96 were identified for the traits i.e., 100-grain weight, harvest index, grain yield, canopy temperature depression and cell membrane thermostability related to heat tolerance under heat stress condition. Two RILs 22 and 40 showing better performance over the environment and year. Twenty-six, out of 81 SSR markers used were found polymorphic in parental genotypes and in RILs. Composite interval mapping (CIM) identified quantitative trait loci (QTL) with significant phenotypic variation for number of spikelets per spike, spike length and biological yield per plant along with heat susceptibility index (HSI). QTL identified for biological yield per plant reported maximum phenotypic variation (50.0%) followed by spike length (46.37%) and number of spikelets per spike (12.59%).
Open Access
Status and evaluation of herbicide resistance in Rumex dentatus L. and its management in wheat
(CCSHAU, Hisar, 2020-12) Dahiya, Seema; Sewhag, Meena
Herbicide resistance in Rumex dentatus in wheat could be a major sustainability issue, which can put the rice-wheat cropping system (RWCS) in IGP under serious threat. The investigation entitled “Status and evaluation of herbicide resistance in Rumex dentatus L. and its management in wheat” comprising of a wellstructured survey carried out during rabi season of 2018-19 (end of January to February 2019), a pot bioassay study conducted at screen house, CCS HAU Regional Research Station, Karnal during rabi season of 2018-19 and a two year field experiment, carried out during rabi seasons of 2017-18 and 2018-19 at KVK, Panipat. The survey was conducted in three districts (Panipat, Karnal and Kurukshetra) of Haryana. Two blocks were selected from each district and two villages from each block and a total number of 120 farmers were covered from 12 villages involving 10 farmers from each village. The diagnostic survey in three districts of Haryana indicating that the use of metsulfuron-methyl is decreasing over the years from 61.1% in 2016-17 to 23.8% in 2018-19 and that of metribuzin is increasing from 6.49% in 2016-17 to 41.27% in 2018-19. Majority of farmers apply herbicide for control of R. dentatus during 35- 35 DAS. More than 80% farmers use flood jet/ cut nozzle instead of using recommended flat fan nozzle by only 19% of total farmers. Almost all farmers have heard about herbicide resistance (99.9%) and they follow herbicide rotation (85.8%) after an average of 2.04 years. Spraying was done mostly by hired labor (70.3%) but in the presence of farmers. Around 75% farmers do not follow crop rotation. Farmers perceptions about poor efficacy of herbicide were mainly poor quality of herbicide and herbicide resistance. Overall farmers assign first rank to 2,4-D with 1.36 rating and after 2,4-D, metribuzin was the second choice of farmers and assign 1.45 rating. Farmers have assigned 3rd position to clodinafop+ metribuzin with 2.07 rating. Metsulfuron-methyl was reported for not showing any control, so it was placed at last position by farmers with overall 2.96 rating. The numbers of farmers going for second spray was increased from 15.9% in 2016-17 to 55.6% in 2018-19 similarly numbers of farmers going for herbicide > 1 to 2 X dose of herbicide has been increased from 5.6% in 2016-17 to 53.0% in 2018-19. It may be concluded that there is problem with control of R. dentatus particularly by use of metsulfuron-methyl in Haryana. From pot bioassay study, it can be concluded that three biotype of R. dentatus viz.Ujha, Bhagwanpur and Gudha have been found resistance to metsulfuron-methyl at graded doses. None of the biotype was found resistance of carfentrazone-ethyl and 2,4-D. The findings of the study established that problem of resistance in R. dentatus in Haryana. From the field experiment, it can be concluded that pendimethalin (1000 g ha-1), 2,4-D ester (600 g ha-1), 2,4-D Na (1000 g ha-1), carfentrazone+ metsulfuron (25 g ha-1) were found at par to weed free treatment and also found as best treatments for controlling resistance population of R. dentatus and beside these carfentrazone-ethyl (20 g ha-1), 2,4-D amine (750 g ha-1) and metribuzin (210 g ha-1) were also found effective for controlling resistance population of R. dentatus. In nutshell, based on this study it may be concluded that resistance has started developing in R. dentatus against metsulfuron-methyl and 2,4-D, carfentrazone-ethyl, pendimethalin, carfentrazone+ metsulfuron and metribuzin could be used as alternate herbicide with proper spray techniques.

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