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Target-Based Physiological Modulations and Chloroplast Proteome Reveals a Drought Resilient Rootstock in Okra (Abelmoschus esculentus) Genotypes.

ABSTRACT: As climate changes increase, drought stress is becoming a problem for all major horticultural crops; among them is okra (Abelmoschus esculentus). Despite its superior resilience to heat stress and high nutritional content, it is still underutilized in contrast to other vegetable crops. Moreover, the drought-resistant and drought-sensitive genotypes of okra are also not well known and require further exploration to improve their productivity. To investigate this in more detail, we performed comparative physiological and large-scale chloroplast proteomics on drought-stressed genotypes of okra. We evaluated four major genotypes of okra, viz., NS7774, NS7772, Green Gold, and OH3312 for drought resilient rootstock. The physiological modulations demonstrated a significant change by 50-76% in biomass, net-photosynthetic machinery, water transport, and absorption both in early and late stages of drought stress compared to well-watered crops in all genotypes. Maximum oxidative damage due to drought stress was observed for the genotypes NS7772, Green Gold and OH3312 as depicted by H₂O₂ and O₂^- determination. Greater oxidative stress was correlated to lesser antioxidant activity and expression of antioxidant enzymes, such as catalase and ascorbate peroxidase under stress in okra genotypes. The overall photosynthetic pigments, such as total chlorophyll, and total carotenoid content, were also decreased, and stomatal guard cells were disrupted and appeared closed compared to the control for the above three mentioned genotypes, except NS7774. A subsequent tissue-specific proteome analysis of chloroplasts and thylakoids analyzed by BN-PAGE (blue native polyacrylamide gel electrophoresis) revealed either over or under expression of specific proteins, such as ATPase, PSI, PSII core dimer, PSII monomer and ATP synthase. The expression of multiprotein complex proteins, including PSII-core dimer and PSII-core monomer, was slightly higher for the genotype NS7774 when compared to three other genotypes for both 5 and 10 days of drought stress. Further identification of specific proteins obtained in second dimension BN-PAGE provided descriptive detail of seven proteins involved in drought resistance across all genotypes. The identified proteins are majorly involved in photosynthesis under drought stress, suggesting NS7774 as a drought tolerant genotype. Further, the proteomic results were confirmed using Immunoblot by selecting specific protein such as PsaA. Overall, from our physiological modulations and chloroplast proteomics in all genotypes, we summarized NS7774 as a resilient rootstock and the other three genotypes (NS7772, OH3312, and Green Gold) as sensitive ones.

SUBMITTER: Razi K

PROVIDER: S-EPMC8657999 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Okra is grown globally for its nutritional and economic benefits. Okra seeds ensure continuous production of the crop but challenges of poor production, adaptability and management may not allow the seeds to express their full potential. There are two seed production systems in Cameroon; the informal and formal. In Buea, the informal seed system is used by most farmers for seed production/utilization and farmers are reluctant to use hybrid seeds. This study aimed to assess the informal seed system of okra and evaluate the adaptability of seed produce from informal and formal systems in Buea. A survey and a field experiment were carried out. The designs for the survey and field experiment were stratified random sampling and randomized complete block design respectively. Data collection for the survey was done using questionnaires and other data collection instruments, while for the field experiment, data was collected on germination, vegetative growth parameters, incidence and severity of pests / diseases and yield. Data analysis for the survey was done using descriptive statistics, while data from the field experiment was done using a two-way ANOVA test and treatment means compared using the Tukey test at 5% probability. Results from the survey showed that women (60%) were mostly involved in seed production by mass selection from two landraces identified. Preservation of seeds was mostly done with the use of wood ash (58%) and insects were the major postharvest pest (76%). For the field experiment, at 66 DAP, Yellen recorded the highest significant number of leaves (13.417), leaf area (771.4 cm2) and the number of branches (5.64). Clemson spineless recorded the highest significant incidence (89.9%) and severity for pests / diseases while Kirikou and Landrace recorded the least incidence (0.0%) and severity. Kirikou recorded the highest significant yield (6.0 tons/ha), followed by Landrace (5.3 tons/ha). These findings reveal the performance of the landrace and provide reasons why farmers in Buea are reluctant to use hybrid okra seeds. The Landrace should certainly have adaptable genes, coupled with the autogamous nature of okra which encourages inbreeding for homozygous traits, which are dominant in expression compared to heterozygous traits.

Project description:IntroductionThe introgression of genetic material from one species to another through wide hybridization and repeated back-crossing, plays an important role in genetic modification and enriching the cultivated gene-pool with novel genetic variations. Okra (Abelmoschus esculentus [(L.) Moench)] is a popular vegetable crop with high dietary fibre and protein, rich in essential amino acids, lysine and tryptophan. The wild Abelmoschus genepool has many desirable traits like ornamental value, short internodal length, more number of productive branches, extended bearing, perennation tendency, reduced fruit length (more consumer preferred trait), high mucilage content (medicinal value), abiotic stress tolerances such as drought, high temperature and biotic stress resistances such as okra Yellow Vein Mosaic Virus (YVMV) and Enation Leaf Curl Virus (ELCV) diseases. The repeated use of elite breeding lines led to narrowing of the genetic base of the okra crop, one of the major factors attributed to breakdown of resistance/ tolerance to biotic stresses. YVMV and ELCV are the two major diseases, causing significant yield loss in okra. Hence, wide hybridization was attempted to transfer tolerance genes from wild species to the cultivated genepool to widen the genetic base.Material and methodsThe screening of germplasm of wild Abelmoschus species at hotspots led to the identification of tolerant species (Abelmoschus pungens var. mizoramensis, A. enbeepeegeearensis, A. caillei, A. tetraphyllus and A. angulosus var. grandiflorus), which were further used in a wide-hybridization programme to generate interspecific hybrids with the cultivated okra. Presence of pre- and post-zygotic barriers to interspecific geneflow, differences in ploidy levels and genotype specific variations in chromosome numbers led to varying degrees of sterility in F1 plants of interspecific crosses. This was overcome by doubling the chromosome number of interspecific hybrids by applying Colchicine at the seedling stage. The 113 cross derivatives generated comprising amphidiploids in the F1 generation (30), F3 (14), one each in F2 and F4 generations, back cross generation in BC1F2 (03), BC1F3 (25), and BC2F3 (02), crosses between amphidiploids (27), multi-cross combinations (07) and inter-specific cross (between A. sagittifolius × A. moschatus subsp. moschatus) selfed derivatives at F8 generation (03) were characterized in the present study. Besides they were advanced through selfing and backcrossing.Results and discussionThe amphidiploids were found to possess many desirable genes with a considerable magnitude of linkage drag. Majority of the wide cross derivatives had an intermediate fruit morphology and dominance of wild characters viz., hispid fruits, stem, leaves, tough fruit fibre, vigorous perennial growth habit and prolonged flowering and fruiting. The fruit morphology of three BC progenies exhibited a high morphological resemblance to the cultivated okra, confirming successful transfer of useful genes to the cultivated okra genepool. The detailed morphological characteristics of the various combinations of Abelmoschus amphidiploids and the genetic enhancement of the genepool achieved in this process is reported here.

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Target-Based Physiological Modulations and Chloroplast Proteome Reveals a Drought Resilient Rootstock in Okra (Abelmoschus esculentus) Genotypes.

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