Field screening and identification of biochemical indices of pod borer (Helicoverpa armigera) resistance in chickpea mutants.

Chickpea pod borer (CPB) (Helicoverpa armigera) is one of the major pests, causing significant yield losses. The objectives were to screen chickpea mutants for pod borer resistance/tolerance under field conditions and identification of biochemical markers of tolerance. Chickpea mutant CM216-A/15 had highest leaf (25 trichomes/mm2) and stem trichome density (17 trichomes/mm2) with least pod damage at Kallur Kot and highest pod weight per plant (22.8 ± 2.6g) at AZRI. Higher total phenolic contents (TPCs) and antioxidant capacity were detected in tolerant mutants, i.e., CM216-A/15 and CM664/15. TPC was positively associated with pod yield and had negative correlation with pod damage. Mutants CM216-A/15, CM664/15, and CM766/15 depicted the highest resilience to CPB, owing to higher hairiness, better antioxidant defense response, and lower levels of hydrolytic enzymes and sugars. Identified biochemical markers like TPC, total oxidant status, superoxide dismutase, and pigments can be used for screening of CPB-tolerant/resistant mutants.

Investigation of Distinctive Morpho-Physio and Biochemical Alterations in Desi Chickpea at Seedling Stage Under Irrigation, Heat, and Combined Stress.

Global climatic instabilities have become the main reason for drastic yield losses in chickpea. This shift in climate could be a great threat in the future for food security in developing countries. Chickpea production is badly hampered by heat stress coupled with drought stress, and these factors can reduce yields by 40-45%. To mitigate yield losses due these abiotic factors, irrigation supplementation could be the best strategy. The present study aimed to (i) investigate the tolerance response of 9 desi chickpea genotypes against heat stress (H), irrigation (I), and a combination of both (I+H) through morphophysiological and biochemical indices at early growth stage, and (ii) assess yield performance across multiple locations of the country. Results revealed that under irrigation treatment, all genotypes perform well, but the genotypes D-09027 and D-09013 showed best performance because, as compared to control, they retained root length, seedling fresh weight, root fresh weight, root dry weight, esterase activity, Malondialdehyde (MDA) content, total chlorophyll, and total carotenoids. Shoot length and total phenolic contents (TPC) increased in both genotypes. Superoxide dismutase (SOD) and peroxidase (POD) increased in D-09027 and retained in D-09013. Catalase activity increased in D-09013 and retained in D-09027. Protease activity, total water potential and osmotic potential decreased in both genotypes and depicted high yield potential with 27 and 30% increase in yield over Bhakhar-2011 (check), respectively. In case of heat stress, maximum tolerance was found in genotypes CH104/06 and D-09013 with no change in shoot and root length, seedling dry weight, shoot fresh and dry weight, root dry weight, relative water content, turgor water potential, catalase (CAT) activity, esterase activity, increased root fresh weight, peroxidase activity (POD), ascorbate peroxidase activity (APX), and lycopene with low accumulation of protease and Malondialdehyde content (MDA). Both genotypes depicted high yield potential with 30 and 43% increase in yield over check across multiple locations of the country. Under the combined treatment, most genotypes showed good performance, while CH104/06 was selected as best performer genotype because significant of its increased root fresh weight, lycopene content, chlorophyll b, total carotenoids, total chlorophyll, retained shoot length, root length, seedling fresh and dry weight, total water potential, osmotic potential, relative water content, peroxidase activity (POD), catalase, esterase, and its ascorbate peroxidase (APX) activity and total soluble proteins (TSP) showed highest yield potential with 43% increase over check. Identified best performing and tolerant genotypes can further be employed for breeding climate-smart chickpea genotypes for sustainable production under changing climate.

Biochemical Profiling for Antioxidant and Therapeutic Potential of Pakistani Chickpea (Cicer arietinum L.) Genetic Resource.

In Pakistan, chickpeas (Cicer arietinum L.) are the largest grown legume crops, especially in desert areas. Along with an excellent source of nutrition, chickpea seeds have discernible medicinal and antioxidant characteristics. The diverse set of 90 chickpea genotypes (66 desi and 24 kabuli) were collected from different research zones in Pakistan, and seed flour was used for biochemical profiling. Genotypes were significantly different (Tukey HSD test, P < 0.05) for the traits under investigation. In non-enzymatic antioxidants, highest seed total phenolic contents (TPC) (34725 ± 275 µM/g s. wt.) was found in CM-98 (desi), ascorbic acid (AsA) (69.23 ± 2.25 µg/g s. wt.) in WH-3 (desi), and total flavonoid content (TFC) (394.98 ± 13.06 µg/mL sample) was detected in WH-11 (desi). In the class of enzymatic antioxidants, the highest seed ascorbate peroxidase (APX) (1680 ± 40 Units/g s. wt.) was detected in Tamman-2013 (kabuli), peroxidases (POD) (2564.10 ± 233.10 Units/g s. wt.) activity in CM1235/08 (desi), and superoxide dismutase (SOD) (279.76 ± 50 Units/g s. wt.) was detected in CH24/11 (desi). Highest seed catalase activity (CAT) (893 ± 50 Units/g s. wt.) and proline content (272.50 ± 20.82 µg/g s. wt.) was detected in an ICC-4951 (desi). In hydrolytic enzymes, the highest activity of esterase (37.05 µM/min/g s. wt) was found in, CH56/09(Kabuli), protease (11080 ± 10 Units/g s. wt.) in Karak-2 (desi), and α-amylase (213.02 ± 3.20 mg/g s. wt.) was observed in CH74/08 (kabuli). In other biochemical parameters, the highest seed total oxidant status (TOS) (356 ± 17.50 µM/g s. wt.) was detected in CM3457/91 (desi); malondialdehyde (MDA) content (295.74 ± 3.097 uM/g s. wt.) was observed in CM-2008 (kabuli), and total antioxidant capacity (TAC) (8.36 ± 0.082 µM/g s. wt.) was found in CM-72 (desi). In case of pigment analysis, Sheenghar-2000 (desi) depicted highest lycopene (12.579 ± 0.313 µg/g s. wt.) and total carotenoids (58.430.23 ± 0.569 µg/g s. wt.) contents. For seed therapeutic potential, the highest seed α-amylase inhibition (82.33 ± 8.06%) was observed in CM-88 (desi), while WH-1, WH-6, and ICCV-96030 (desi) depicted the highest value for seed anti-inflammatory potential (78.88 ± 0.55%). Genotypes with the highest antioxidant and therapeutic potential can be utilized as a natural antioxidant source and in breeding programs aimed at improving these traits in new breeding lines.

Selection and screening of drought tolerant high yielding chickpea genotypes based on physio-biochemical indices and multi-environmental yield trials.

BACKGROUND: Chickpea is one of the major legume crops being cultivated in the arid and semi-arid regions of Pakistan. It is mainly grown on the marginal areas where, terminal drought stress is one of the serious threats to its productivity. For defining the appropriate selection criteria for screening drought tolerant chickpea genotypes, present study was conducted. Distinct chickpea germplasm was collected from different pulses breeding institutes of Pakistan and evaluated for drought tolerance at germination and early seedling stages, furthermore, at late vegetative growth stages physiochemical traits and multi-environment yield performance were also tested. RESULTS: Chickpea genotypes under different environments, were significantly varied for different seedling traits, physio-chemical attributes and seed yield. Genotypes showing drought tolerance by performing better at an early seedling stages were not correspondingly high yielding. Screening for drought tolerance on seed yield basis is the most appropriate trait to develop the drought tolerant as well as high yielding chickpea genotypes. Results confirmed that traits of early growth stages were not reflecting the drought tolerance at terminal growth stages and also did not confer high yielding. NIAB-rain fed environment proved ideal in nature to screen the chickpea genotypes whereas, NIAB-lysimeter and Kalur Kot was least effective for selecting genotypes with high seed yield. Genotypes D0091-10, K010-10, D0085-10, K005-10, D0078-10, 08AG016, 08AG004, D0080-10, 09AG002, K002-10 and D0099-10 were high yielding and drought tolerant based on their performance across multiple hotspot environments. CONCLUSIONS: The selected genotypes are intended for further evaluation for varietal approval to recommend for general cultivation on farmer fields in drought hit areas of Pakistan. Among physio-biochemical traits, higher proline, glycine betain, RWC and CMS were reflecting the higher capability to tolerate the drought stress in chickpea. Drought sensitive genotypes (K0037-10, 2204, K0052-10, 09AG015, K0042-10, CM709/06, K0068-10, K004-10, K0026-10 and K0063-10) were also identified in present study which were resourceful asset for using as contrasting parents in hybridization programs. To our knowledge, this is first report using an integrated approach involving, physio-biochemical indices, and multi-environmental yield trials, for comparison, screening and selection of chickpea genotypes for drought tolerance.

A Comparative Study for Assessing the Drought-Tolerance of Chickpea Under Varying Natural Growth Environments.

This study was planned with the purpose of evaluating the drought tolerance of advanced breeding lines of chickpea in natural field conditions. Two methods were employed to impose field conditions; the first: simulating drought stress by growing chickpea genotypes at five rainfed areas, with Faisalabad as the non-stressed control environment; and the second: planting chickpea genotypes in spring to simulate a drought stress environment, with winter-sowing serving as the non-stressed environment. Additive main effects and multiplicative interaction (AMMI) and generalized linear models (GLM) models were both found to be equally effective in extracting main effects in the rainfed experiment. Results demonstrated that environment influenced seed yield, number of primary and secondary branches, number of pods, and number of seeds most predominantly; however, genotype was the main source of variation in 100 seed weight and plant height. The GGE biplot showed that Faisalabad, Kallur Kot, and Bhakkar were contributing the most in the GEI, respectively, while Bahawalpur, Bhawana, and Karor were relatively stable environments, respectively. Faisalabad was the most, and Bhakkar the least productive in terms of seed yield. The best genotypes to grow in non-stressed environments were CH39/08, CH40/09, and CH15/11, whereas CH28/07 and CH39/08 were found suitable for both conditions. CH55/09 displayed the best performance in stress conditions only. The AMMI stability and drought-tolerance indices enabled us to select genotypes with differential performance in both conditions. It is therefore concluded that the spring-sown experiment revealed a high-grade drought stress imposition on plants, and that the genotypes selected by both methods shared quite similar rankings, and also that manually computed drought-tolerance indices are also comparable for usage for better genotypic selections. This study could provide sufficient evidence for using the aforementioned as drought-tolerance evaluation methods, especially for countries and research organizations who have limited resources and funding for conducting multilocation trials, and performing sophisticated analyses on expensive software.

Pre-breeding of lentil (Lens culinaris Medik.) for herbicide resistance through seed mutagenesis.

Lentil is a poor competitor of weeds and its sensitivity to herbicides is a major hurdle for large scale production. The present study was conducted to select herbicide resistant lentil genotypes through seed mutagenesis. Seeds of three advanced lentil genotypes (LPP 11001, LPP 11100 and LPP 11116) were treated with two different concentrations of ethyl methanesulfonate (EMS; 0.1 and 0.2%), hydrazine hydrate (HH; 0.02 and 0.03%) and sodium azide (SA; 0.01 and 0.02%) to develop M1 seed. The M2 was screened against two herbicides including Ally Max 28.6% SG (X = 34.58 g/ha and 1.5X = 51.87 g/ha) and Atlantis 3.6% WG (X = 395.2 g/ha and 1.5X = 592.8 g/ha) using the following three screening methods: post plant emergence (PPE), pre-plant incorporation (PPI) and seed priming (SP). Data were recorded on survival index and survival percentage from each experimental unit of every population. Plants in all populations were categorized following their reaction to herbicides. The newly developed populations showed greater variation for herbicide resistance when compared to their progenitors. Phenotypic traits were significantly reduced in all the screening environments. Overall, 671 herbicide resistant mutants were selected from all testing environments. The seeds from selected plants were re-mutagenized at 150 Gy of gamma radiation and evaluated against higher dose of herbicides. This allowed selection of 134 herbicide resistant mutants. The selected mutants are useful germplasm for herbicide resistance breeding of lentil.

Association of plant growth-promoting Serratia spp. with the root nodules of chickpea.

Serratia species-affiliated DNA sequences have recently been discovered in the root nodules of two chickpea cultivars; however, little is known about their potential influence on chickpea plant growth. All Serratia-affiliated sequences (1136) could be grouped into two clusters at 98% DNA similarity. The major cluster, represented by 96% of sequences, was closely associated with Serratia marcescens sequences from GenBank. In the current study, we isolated two Serratia strains, 5D and RTL100, from root nodules of a field-grown Desi cultivar from Faisalabad and Thal areas, respectively. In vitro, strain 5D showed significantly higher phosphate (P) solubilization and lactic acid production than RTL100, whereas a comparable concentration of phytohormone was produced by both isolates. The application of Serratia strain 5D as an inoculum resulted in 25.55% and 30.85% increases in the grain yield of crops grown on fertile soil in irrigated areas and nutrient-deficient soil in rainfed areas, respectively, compared to the non-inoculated control. Results of plant inoculations indicated that Serratia sp. 5D and RTL100 can serve as effective microbial inoculants, particularly in nutrient-deficient soils in rainfed areas, where chickpea is the only major crop grown during the entire year.

Detection of quantitative trait loci for mungbean yellow mosaic India virus (MYMIV) resistance in mungbean (Vigna radiata (L.) Wilczek) in India and Pakistan.

Yellow mosaic disease (YMD) is one of the major diseases affecting mungbean (Vigna radiata (L.) Wilczek). In this study, we report the mapping of the quantitative trait locus (QTL) for mungbean yellow mosaic India virus (MYMIV) resistance in mungbean. An F8 recombinant inbred line (RIL) mapping population was generated in Thailand from a cross between NM10-12-1 (MYMIV resistance) and KPS2 (MYMIV susceptible). One hundred and twenty-two RILs and their parents were evaluated for MYMIV resistance in infested fields in India and Pakistan. A genetic linkage map was developed for the RIL population using simple sequence repeat (SSR) markers. Composite interval mapping identified five QTLs for MYMIV resistance: three QTLs for India (qYMIV1, qYMIV2 and qYMIV3) and two QTLs for Pakistan (qYMIV4 and qYMIV5). qYMIV1, qYMIV2, qYMIV3, qYMIV4 and qYMIV5 explained 9.33%, 10.61%, 12.55%, 21.93% and 6.24% of variation in disease responses, respectively. qYMIV1 and qYMIV4 appeared to be the same locus and were common to a major QTL for MYMIV resistance in India identified previously using a different resistant mungbean.