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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
Mapping and Monitoring of Agroforestry areas in Yamunanagar, Ambala and Panchkula districts of Haryana state using Geospatial techniques
(CCS HAU, Hisar, 2021-07-30) Bojja Harish Babu; Bimlendra Kumari
The study was conducted to map and monitor the agroforestry areas in Yamunanagar, Ambala and Panchkula districts of Haryana state from the year 2000 to 2019 through geospatial techniques using LISS-III (2000, 2005 and 2010) and LISS-IV (2015 and 2019) satellite data. Apart from mapping and monitoring, an attempt has been made to identify potential agroforestry sites like scrubs, fallows and piedmont areas on LISS-IV dataset of 2019 by on screen digitization technique for future cultivation of agroforestry species by farmers in the study area. Supervised method of classification technique was adopted to classify LU/LC classes like agroforestry, agriculture, forest, built-up, waterbodies, plantations and riverbed areas. The results of the study revealed that, agroforestry area delineated in Yamunanagar for the year 2000 (3,898.62 ha), 2005 (5,403.17 ha), 2010 (7,134.90 ha), 2015 (14,650.51 ha) and 2019 (17,022.76 ha). In Ambala 2000 (2,696.48 ha), 2005 (3,745.33 ha), 2010 (7,400.92 ha), 2015 (12,350.91 ha) and 2019 (15,552.53 ha). In Panchkula 2000 (1,098.15 ha), 2005 (1,212.8 ha), 2010 (1,784.03 ha), 2015 (2,034.87 ha) and 2019 (2,367.12 ha). The accuracy of the image classification was delineated based on the ground truth data and accuracy found in agroforestry was 90% for Yamunanagar, 95% for Ambala and 93% for Panchkula districts By the extraction of vector layers from the classified raster image, blockwise monitoring of agroforestry areas was delineated and change in various categories of LU/LC by performing overlay analysis in ArcGIS platform was detected and results revealed were (5,678.5 ha) in Yamunanagar, (6,783.6 ha) in Ambala and (1,202.6 ha) in Panchkula, the agriculture areas were converted to agroforestry areas from the year 2000 to 2010 and (14,486.6 ha) in Yamunanagar, (13,534.8 ha) in Ambala and (1,866.2 ha) in Panchkula, the agriculture areas were converted to agroforestry areas from the year 2010 to 2019. Based on the groundwater quality, soil condition and rainfall factor in the study area, potential sites were digitized, the results revealed that, about 10,525.76 ha of the potential sites were identified and mapped in Yamunanagar, around 12,179. 85 ha in Ambala and 4,023 ha in Panchkula districts. Based on the study, it is concluded that with the use of LISS-III and LISS-IV satellite data accurate mapping of agroforestry areas can be delineated which would help the planners in monitoring and identification of hotspots for suitable interventions in the districts.
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Genome-wide association mapping in barley for terminal heat tolerance and malting quality
(CCSHAU, Hisar, 2021-07) Verma, Swati; Yashveer, Shikha
Barley (Hordeum vulgare L.), is one of the major cereal crops and has been cultivated since ancient times in different parts of the world. In particular, heat stress at the post-heading stage causes considerable yield reduction due to the stress. Understanding the genetic variation, changes in physiological processes, and level of genetic diversity existing among genotypes are needed to produce new cultivars not only having a high tolerance to heat stress, but also displaying high yield. So to address this challenge, a panel of 316 diverse barley genotypes (AM2017) were procured. It was evaluated under two conditions timely-sown (TS) and late-sown (LS) conditions in two seasons of 2017-18 and 2018-19 at CCSHAU, Hisar. Ten agro-morphological, four physiological, and five grain malting quality traits were studied. Genetic diversity and population structure were explored in the AM2017 panel after genotyping it with the 50 K iSelect Illumina Barley SNP array. A set of 36,793 SNP markers, covering a genetic distance of 991.82 cM with an average marker density of 37.09 SNPs/cM, was obtained after quality filtration. The gene diversity (GD) and Polymorphic Information Content (PIC) at the genome level were 0.362 and 0.289, respectively. The phenotypic results showed a clear reduction in trait performance under the LS condition. Marker-trait associations (MTAs) were estimated using the compressed mixed linear model. Based on the estimated MTAs and linkage disequilibrium (LD) decay observed in the genome, putative QTL was identified as associated with the traits studied. The most robust QTL observed under both sowing conditions were further studied and validated using the previously reported QTL. Several traits were found to have QTL that was not found to be reported. The combination of analyses using SNPs and changes in physiological traits provided useful information on genomic regions taking part in heat stress tolerance.
Unknown
Performance of wheat under different irrigation and nitrogen sources
(CCSHAU, Hisar, 2021-10) Vikram Singh; Parveen Kumar
The present investigation was carried out at Vegetable Research Farm, CCS HAU, Haryana to evaluate the performance of wheat under various irrigation and nitrogen sources. The trial was conducted in split-plot design with three replication. It consisted of three irrigation sources i.e. treated sewage water (TSW), canal water (CW) and tube well water (TW); and seven nitrogen sources i.e. control (T1), 100 % RDN through chemical fertilizers (T2), 100 % RDN through vermicompost (T3), 50 % RDN through chemical fertilizers + 50 % RDN through vermicompost (T4), 100 % RDN through FYM (T5), 50 % RDN through chemical fertilizers + 50 % RDN through FYM (T6), 50 % RDN through chemical fertilizers +25 % RDN through FYM +25 % RDN through vermicompost (T7). The organic manures was applied before sowing of crop. The results revealed that the plant population at 15 DAS did not differ significantly among various irrigation and nitrogen sources during both the kharif seasons. Maximum plant height (cm), dry matter accumulation (g/plant) and growth indices viz. leaf area index, leaf area duration, crop growth rate and chlorophyll content; and yield attributing characters i.e. number of effective tillers/m2, spike length, number of grains per spike and test weight, and yields i.e. grain, straw and biological yield were recorded with treated sewage water over canal and tubewell water. However, among nitrogen sources, the significantly highest growth parameters and indices, and yields were obtained under application of 100 % RDN through chemical fertilizer (T2) was at par with 50 % RDN through fertilizers + 50 % RDN through vermicompost (T4), 50 % RDN through fertilizers + 25 % RDN through FYM + 25 % RDN through vermicompost (T7) and 50 % RDN through fertilizers + 50 % RDN through FYM (T6).The application of various irrigation and nitrogen sources gave non-significantly relative growth rate (mg/g/day) of wheat during both the years (2017-19). Different irrigation treatments were failed to produce any significant effect on harvest index during both the growing seasons, while the treatments receiving chemical fertilizer application alone or in combination with organic manures (vermicompost and FYM) gave at par harvest and attraction index over control. The significantly improved wheat grain quality in terms of protein content, protein yield, sedimentation value and hectolitre weight was observed under application of treated sewage water which was statistically at par with canal water. However, among nitrogen sources, the application of 100% RDN through chemical fertilizers improved the grain quality but it was at par with 50 % RDN through fertilizers + 50 % RDN through vermicompost (T4), 50 % RDN through fertilizers + 25 % RDN through FYM + 25 % RDN through vermicompost (T7) and 50 % RDN through fertilizers + 50 % RDN through FYM (T6).The treatments receiving treated sewage water gave higher nutrients (N, P, and K) concentration and their uptake in grain and straws of wheat crop during both the years. While among the nitrogen sources, treatment T2 exhibited higher nutrient accumulation over rest of the treatments. However, T2 remained at par with T4, T7 and T6. The effects of irrigation sources were non-significant on water consumptive use during both the years. It was observed that plants irrigated with treated sewage water used the water more efficiently compared to canal and tube well water. However, the effect of irrigation sources was non-significant during year 2017-18. Among nitrogen sources, the application of organic manures alone or in combination with chemical fertilizers gave comparable water use efficiency over control due to increased grain yield. During both years of the experimentation, the highest economic return regards net return and B: C ratio were attained with application of treated sewage water. Among fertilizer treatments, the highest B: C was recorded under treatments where sole application of chemical fertilizers were applied. However, integrated application of chemical fertilizers and organic manures gave higher return over sole application of manures and control. Compared to the initial soil pH values, it was reduced under use of all type of irrigation water. However, the application of treated sewage water significantly reduced soil pH as compared to canal and tubewell water. As the EC of tubewell water is high, so after two years of wheat cultivation, the soil EC was significantly increased from 0.84 to 0.91 dS/m. The build-up of soil fertility status regards was organic carbon, available N, P and K was noticed under treated sewage water irrigated plots. However, among different nitrogen sources, the significant improvement in organic carbon, available N, P and K in the soil was brought be the addition of organic manures i.e. 50 % RDN through vermicompost and farm yard manures over sole application of RDN through mineral fertilizers and control treatments. Hence, use of sewage water under wheat cultivation with integrated nitrogen application of chemical fertilizers and organic manures proved economically beneficial.
Open Access
Effect of residue retention and tillage practices on performance of wheat varieties
(CCSHAU, Hisar, 2021-06) Malik, Kapil; Lathwal, O. P
The present study entitled, “Effect of residue retention and tillage practices on performance of wheat varieties” was conducted at the farm of college of agriculture, Kaul during rabi seasons of 2017-18 and 2018-19. The experiment was laid out in strip plot design, keeping four sowing methods (wheat sown with turbo seeder under full residue retention, TS+IR+LR; wheat sown with turbo seeder under intact rice residue, TS+IR; wheat sown with zero till seed cum fertilizer drill under no residue, ZT+NR; and wheat sown in conventional tillage conditions under no residue, CT+NR) in main plots and six wheat varieties (HD 2967, HD 3086, WH 1105, WH 711, WH 1124 and WH 1142) in sub plots with three replications. Among sowing methods during both the years, TS+IR+LR took significantly maximum number of days to attain different phenological stages, recorded better growth, yield and quality attributes and resulted into significantly higher wheat yields as compared to CT+NR. The net returns benefit cost ratio and nutrient uptake were also maximum with TS + IR + LR sowing method. After completion of the experiment, soils under TS+IR+LR registered significantly higher nitrogen content than under CT+NR. Among the wheat varieties, HD 3086 recorded maximum growth parameters (dry matter, crop growth rate, leaf area index and number of tillers), yield attributes (effective tillers and test weight) and grain yield but was strictly followed by WH 1105 and HD 2967 and significantly higher than rest of varieties. Quality parameters viz; hectolitre weight and grain appearance score were also higher in HD 3086. However, spike length, number of grains/spike, protein content, and nitrogen content in seed, straw yield and biological yield were found maximum in wheat variety WH 1105. Variety WH 1142 recorded maximum plant height and required maximum days to attain different phenological stages than rest of varieties. The emergence count, phosphorus content, available P and K were not differed among wheat varieties. Net returns and benefit cost ratio were obtained maximum with HD 3086 followed by WH 1105, HD 2967, WH 1142, WH 711 and WH 1124 wheat varieties.
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).
Unknown
Applicability of salicylic acid and glycinebetaine to alleviate the effect of salinity stress in Indian Mustard (Brassica juncea (L.) Czern and Coss)
(CCSHAU, Hisar, 2021-06) Agarwal, Vishnu Prakash; Neeraj Kumar
The present investigation was carried out to study the effect of salicylic acid (SA) and glycinebetaine (GB) to mitigate the adverse effect of salinity on Indian mustard by growing seeds of genotype RH 0725 in plastic pots filled with dune sand maintaining four different levels of salinity (control, 8, 10 and 12 dS m-1). The crop was sprayed with different concentrations of SA (control, 0.5 mM, 1.0 mM and 1.5 mM) and GB (control, 10 mM, 20 mM and 30 mM) at 40 and 60 days after sowing (DAS). Sampling was done at five days after spray i.e. 45 and 65 DAS. The growth and reproductive parameters, physio-biochemical attributes, antioxidant defense mechanism, yield and yield attributing characters, and protein profile through SDS-PAGE were observed at various growth stages of the crop. Results showed that photosynthetic rate, transpiration rate, stomatal conductance, relative water content (RWC), water potential (Ψw), osmotic potential (Ψs), quantum yield, chlorophyll content (SPAD), chlorophyll stability index (CSI), pollen viability, pollen germination and tube growth, days to maturity, plant height, number of branches (plant-1), number of siliqua (plant-1), number of seeds (siliquae-1), test weight (g), biological yield (BY; g plant-1), economical yield (EY; g plant-1) and salt susceptibility index (SSI); protein, ascorbate oxidized (DHA) and glutathione oxidized (GSSG) contents were decreased whereas days to 50 % flowering (DTA), proline, glycine-betaine, total soluble sugar (TSS), hydrogen peroxide (H2O2) and malondialdehyde (MDA) contents; relative stress injury (RSI), specific activity of antioxidants; and ascorbate reduced (AsA) and glutathione reduced (GSH) contents were increased with increasing salinity levels. Foliar application of SA (upto 1.0 mM) and GB (upto 20 mM) significantly increased the days to maturity, pollen viability, pollen germination and tube growth, photosynthetic rate and stomatal conductance, RWC, Ψw, quantum yield, chlorophyll content (SPAD), CSI; proline, glycine-betaine, TSS and protein contents, specific activity of antioxidants, and AsA, DHA, GSH and GSSG contents in leaves and roots at both the sampling stages (45 and 65 DAS); and all the yield and yield attributes but slight early in DTA and reduced transpiration rate, RSI, and H2O2 and MDA contents over control in all the salinity levels. Application of SA 1.5 mM showed opposite effects on salinity tolerance as observed in SA 1.0 mM treatment. Application of GB 30 mM gradually enhanced the salinity tolerance by improving all the parameters but the rate of improvement was lower than that of GB 20 mM treatment. The SSI was lower recorded in SA 1.0 mM treatment followed by GB 30 mM treatment in all the salinity levels. The correlation coefficient analysis of various morpho-physiological, biochemical and yield characters indicating the contribution in imparting EY of Indian mustard. Analysis of protein samples in SDS-PAGE further confirmed the results. Overall, it is concluded that all the morpho-physiological, biochemical, yield and yield attributes of Indian mustard genotype (RH 0725) were decreased on account of increasing salinity whereas DTA, proline, glycine-betaine, TSS, H2O2, MDA, RSI, specific activity of antioxidants and, AsA and GSH contents were increased. Sprays of SA 1.0 mM and GB 20 mM particularly at 40 and 60 DAS showed

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