Mutations induced in some Egyptian cowpea varieties with yield characteristics and high nutritional value using gamma rays and evaluation by microsatellite markers.

PURPOSE: In order to achieve mutations with enhanced economic, productive, and nutritional characteristics in the two Egyptian cowpea varieties, Dokki 331 and Kaha 1, the application of gamma irradiation at different doses is employed. Additionally, this method aids in distinguishing between these mutations using simple sequence repeat (SSR) analysis. MATERIALS AND METHODS: Two different cowpea cultivars were subjected to varying doses of gamma radiation ranging from 50 to 300 Gy. In order to analyze the effects of radiation, both unirradiated and irradiated seeds from both cultivars were planted using a randomized complete block design. This experiment was conducted over a span of six generations, namely M1, M2, M3, M4, M5, and M6, starting from April 2017 and continuing until 2022. Among the various radiation doses, the cultivar Kaha 1 produced promising traits when exposed to a dose of 150 Gy, while the cultivar Dokki 331 showed favorable traits when exposed to a dose of 300 Gy. These traits were further cultivated and studied until the M6 generation. RESULTS: Induced mutations in two Egyptian cowpea varieties, Kaha 1 and Dokki 331, are subjected to varying doses of gamma radiation (0, 50, 100, 150, 200, 250, and 300 Gy). Morphological and genetic variations were observed, with mutations being induced at doses of 150 Gy for Kaha 1 and 300 Gy for Dokki 331. The mutation in Kaha 1 (beam 1) resulted in dwarfism, altered leaf shape, early flowering, increased peduncles, pods, and pod seed numbers, ultimately leading to enhanced seed production and acreage productivity. In Dokki 331, the mutations primarily affected pod color, resulting in greenish-brown pods with mosaic seeds, segregating black and gray seeds from the mosaic ones. These mutations led to an increase in the nutritional value of the seeds, including higher nitrogen content, total free amino acids, crude protein, total carbohydrates, and total sugars. The genetic diversity of the seven cowpea mutations was assessed using 20 microsatellite markers. The analysis revealed a total of 60 alleles, with an average of three alleles per locus. The allele frequency ranged from 0.2857 to 1.0, with an average of 0.6036. Gene diversity varied from 0.0 to 0.8163, while the heterozygosity was mostly zero, except for one primer (VM 37) with an average of 0.0071. The polymorphic information content (PIC) ranged from 0.7913 to 0.0, with an average of 0.4323. The Marker Index value ranged from 0.36 to 0.0, with an average of 0.152. Overall, our findings demonstrate the successful induction of mutations in Egyptian cowpea varieties using gamma rays, resulting in improved yield characteristics and nutritional value. CONCLUSIONS: Radiation as a physical mutagen is highly regarded for its effectiveness, affordability, speed, and safety in inducing mutations. Utilizing gamma rays, we successfully derived a novel cowpea variety called beam 1 mutation, which has gained approval from the Egyptian Ministry of Agriculture.

Gamma-rays induced genome wide stable mutations in cowpea deciphered through whole genome sequencing.

PURPOSE: Gamma rays are the most widely exploited physical mutagen in plant mutation breeding. They are known to be involved in the development of more than 60% of global cowpea (Vigna unguiculata (L.) Walp.) mutant varieties. Nevertheless, the nature and type of genome-wide mutations induced by gamma rays have not been studied in cowpea and therefore, the present investigation was undertaken. MATERIALS AND METHODS: Genomic DNAs from three stable gamma rays-induced mutants (large seed size, small seed size and disease resistant mutant) of cowpea cultivar 'CPD103' in M6 generation along with its progenitor were used for Illumina-based whole-genome resequencing. RESULTS: Gamma rays induced a relatively higher frequency (88.9%) of single base substitutions (SBSs) with an average transition to transversion ratio (Ti/Tv) of 3.51 in M6 generation. A > G transitions, including its complementary T > C transitions, predominated the transition mutations, while all four types of transversion mutations were detected with frequencies over 6.5%. Indels (small insertions and deletions) constituted about 11% of the total induced variations, wherein small insertions (6.3%) were relatively more prominent than small deletions (4.8%). Among the indels, single-base indels and, in particular, those involving A/T bases showed a preponderance, albeit indels of up to three bases were detected in low proportions. Distributed across all 11 chromosomes, only a fraction of SBSs (19.45%) and indels (20.2%) potentially altered the encoded amino acids/peptides. The inherent mutation rate induced by gamma rays in cowpea was observed to be in the order of 1.4 × 10-7 per base pair in M6 generation. CONCLUSION: Gamma-rays with a greater tendency to induce SBSs and, to a lesser extent, indels could be efficiently and effectively exploited in cowpea mutation breeding.

Brassinolide soaking and preharvest UV-B radiation influence the shelf life of small black bean sprouts.

This study explored the effects of brassinolide (BR) soaking, preharvest ultraviolet-B (UV-B) radiation, and their combined treatments on physiological characteristics, chlorophyll fluorescence, and quality of small black bean sprouts during storage. Results indicated that the combined treatments significantly enhanced contents of flavone, free amino acid, and photosynthetic pigment, and activities of phenylalanine ammonia lyase (PAL) and 2-diphenyl-1-picrylhydrazyl(DPPH) radical scavenging in sprouts stored for 5 days compared with BR treatment alone. The combined treatments significantly increased total phenols content and PAL activity, and reduced malonaldehyde content in sprouts compared with UV-B radiation alone. The inhibitory effect of BR or UV-B on fluorescence of photosystem II was weakened by their combined treatments. Comprehensive analysis indicated that the combined treatments could be used to maintain postharvest small black bean sprouts with high levels of nutritional ingredients by probably keeping high photosynthetic capacity, PAL activity, and DPPH radical scavenging rate in sprouts.

Characterization of 60CO γ-ray induced pod trait of blackgram-A promising yield mutants.

Purpose: Blackgram (Vigna mungo (L.) Hepper) possesses natural flower dropping character with self-pollination, which was highly challenging to create variability through hybridization. Gamma irradiation is a powerful tool to induce genetic alteration and improvement in crops with beneficial mutants. Hence, the present investigation focused on finding out the mutant characters related to the improvement of the agronomic characters and yield-related traits by using gamma irradiation. Pod mutant isolated with fruitful agronomic trait at M2 (Mutagenic) generation was identified and isolated by gamma irradiation is a central focus in the present study, which enhances the yield of the blackgram.Materials and methods: Viable and uniform size of blackgram Vamban-4 seeds were irradiated at a different dose of 60CO (Cobalt) γ-rays viz., 200, 400, 600, 800, 1000, and 1200 Gray (Gy). The further generations were screened and raised to M3 and M4 plants to select the stable mutants with yield traits.Results: In the M2 generation, the following viable and pod mutants were identified, such as tall, dwarf, multipod, small pod, bold pod, striata pod. Morphological traits, such as plant height (PH), number of branches per plant (NBP), number of leaves per plant (NLP), number of cluster per plant (NCP), number of pods per plant (NPP), number of seeds per pod (NSP), pod length (PL), a hundred seed weight (HSW), seed yield per plant (SYP), were measured. Among the mutants, the increased morphogenetic traits were recorded in tall mutants (600 Gy), and the enhancement of yield characteristics noticed in multipod (800 Gy) and bold pod mutants (600 Gy). Compared to other doses, 600 Gy shows enhanced quantitative traits for blackgram improvement.Conclusion: The 60CO γ-rays induced genetic variation in the yield and its related traits and obtained pod mutants, which are useful to enhance the commercial value of Vigna species and crop improvement.

Assessment irradiation effects on different development stages of Callosobruchus maculatus and on chemical, physical and microbiological quality of cowpea seeds.

Irradiation is a physical, environmentally friendly treatment which does not leave any residues in the product. It is increasingly used as an alternative to methyl bromide and other chemical fumigants for disinfestation of insect pest in stored grains. In this research, we try to evaluate the effect of low gamma irradiation doses in the range of 50-1200 Gy on the different stages of Callosobruchus maculatus. It was found that no adults emerged after irradiation of eggs at 450 Gy. No emerging adults (F1 generation) have been shown when larvae or pupae were exposed to 650 Gy. Also, no emerging adult has been shown from the parental C. maculatus adults irradiated with 650 Gy. When suppression of F1 generation was used for the measuring effective irradiation dose and phytosanitary efficacy, 650 Gy was required for disinfestation of cowpea seed weevils. The irradiation dose level of 650 Gy was used for the large-scale confirmatory tests applied to 27,754 adults of C. maculatus in cowpea seeds resulting in non F1 adults' production with a confidence level of 93.77%. All the physical and chemical characteristics of cowpea seeds were non-significantly (P ≤ 0.05) affected by the irradiation dose of 650 Gy. Meanwhile, this irradiation dose very slightly reduced the microbial load of cowpea seeds. We recommend the generic dose 650 Gy as the phytosanitary irradiation dose for the cowpea seed weevil.

Irradiation of mung beans (Vigna radiata): A prospective study correlating the properties of starch and grains.

In this work, the effect of Gamma-irradiation was evaluated on the characteristics of mung bean (Vigna radiata) grains and starches, considering doses up to 5 kGy. For this purpose, the starch structure and properties were evaluated, as well as the grains' hydration, germination and cooking. The irradiation process was able to change the characteristics of both mung bean starches and grains. The starch structure was partially changed, presenting smaller molecules and small changes in the granule morphology. No alterations were observed in the starch X-ray diffraction pattern, while lower pH was achieved. Considering the starch properties, it was observed lower water retention ability at 75 °C, lower apparent viscosity, higher paste clarity and, in general, harder and less viscous gels. The ionizing radiation accelerated the hydration, reduced the germination capacity and improved cooking time of the mung bean grains. The results proved the efficacy of using ionizing radiation, at the doses applied in this work, to desirably modify the mung bean starch and grains.

Impact of infrared treatment on quality and fungal decontamination of mung bean (Vigna radiata L.) inoculated with Aspergillus spp.

BACKGROUND: Mung bean is a rich source of protein, carbohydrates and fiber content. It also exhibits a high level of antioxidant activity due to the presence of phenolic compounds. Aspergillus flavus and A. niger are the two major fungal strains associated with stored mung bean that lead to post-harvest losses of grains and also cause serious health risks to human beings. Thus there is a need to explore an economical decontamination method that can be used without affecting the biochemical parameters of grains. RESULTS: It was observed that infrared (IR) treatment of mung bean surface up to 70 °C for 5 min at an intensity of 0.299 kW m-2 led to complete visible inhibition of fungal growth. Scanning electron microscopy revealed that surface irregularities and physical disruption of spores coat are the major reasons behind the inactivation of IR-treated fungal spores. It was also reported that IR treatment up to 70 °C for 5 min does not cause any negative impact on the biochemical and physical properties of mung bean. CONCLUSION: From the results of the present study, it was concluded that IR treatment at 70 °C for 5 min using an IR source having an intensity of 0.299 kW m-2 can be successfully used as a method of fungal decontamination. The fungal spore population was reduced (approximately 5.3 log10 CFU g-1 reductions) without significantly altering the biochemical and physical properties of grains. © 2017 Society of Chemical Industry.

Effects of gamma-irradiation on cotyledon cell separation and pectin solubilisation in hard-to-cook cowpeas.

BACKGROUND: Cowpeas stored under high temperature and humidity develop the hard-to-cook defect (HTC). This defect greatly increases cooking times and energy costs. To better understand the mechanisms involved in the HTC defect development, the effects of gamma-irradiation on cotyledon cellular structure and pectin solubility in two cowpea cultivars with different susceptibility to HTC defect were investigated. RESULTS: Gamma-irradiation decreased cotyledon cell wall thickness, increased cell size, and intercellular spaces in both cowpea cultivars and reduced cooking time of the less HTC susceptible cultivar. However, it did not reverse the HTC defect in the susceptible cultivar. Gamma-irradiation also increased the levels of cold water- and hot water-soluble pectin. The irradiation effects were thus mainly due to hydrolysis of pectin fractions in the cell walls. However, chelator-soluble pectin (CSP) solubility was not affected. CONCLUSION: As the cell wall changes brought about by gamma-irradiation were associated with pectin solubilisation, this supports the phytate-phytase-pectin theory as a major cause of the HTC defect. However, the non-reversal of the defect in HTC susceptible cowpeas and the absence of an effect on CSP indicate that other mechanisms are involved in HTC defect development in cowpeas, possibly the formation of alkali-soluble, ester bonded pectins. © 2017 Society of Chemical Industry.

Effects of light quality on pod elongation in soybean (Glycine max (L.) Merr.) and cowpea (Vigna unguiculata (L.) Walp.).

Soybean pods are located at the nodes, where they are in the shadow, whereas cowpea pods are located outside of the leaves and are exposed to sunlight. To compare the effects of light quality on pod growth in soybean and cowpea, we measured the length of pods treated with white, blue, red or far-red light. In both species, pods elongated faster during the dark period than during the light period in all light treatments except red light treatment in cowpea. Red light significantly suppressed pod elongation in soybean during the dark and light periods. On the other hand, the elongation of cowpea pods treated with red light markedly promoted during the light period. These results suggested that the difference in the pod set sites between soybean and cowpea might account for the difference in their red light responses for pod growth.

Partial alleviation of oxidative stress induced by gamma irradiation in Vigna radiata by polyamine treatment.

PURPOSE: Environmental changes generate free radicals and reactive oxygen species (ROS) resulting in abiotic stress in plants. This causes alterations in germination, morphology, growth and development ultimately leading to yield loss. Gamma irradiation was used to experimentally induce oxidative damage in an important pulse crop Vigna radiata (L.) Wilczek or mung bean. Our research was aimed towards augmentation of oxidative stress tolerance through treatment with a group of aliphatic amines known as polyamines. MATERIALS AND METHODS: We used sub-lethal doses of gamma irradiation to generate oxidative damage which was evaluated using Nitro blue tetrazolium (NBT) staining, total antioxidant activity, 1, 1-Diphenyl-2-picryl hydrazyl (DPPH) radical scavenging assay, proline content and lipid peroxidation. Changes in internal free polyamines and messenger ribonucleic acid (mRNA) expression of key rate-limiting S-adenosylmethionine decarboxylase (SAMDC) enzyme in polyamine biosynthetic pathway was studied using real-time polymerase chain reaction (PCR). RESULTS: We observed increased oxidative damage with higher irradiation dose which was partially alleviated by putrescine treatment. Internal levels of putrescine and spermidine increased with 1 mM (50 and 100 Gy) and 2 mM putrescine treatment. Expression of SAMDC also increased with putrescine treatment. CONCLUSION: This study shows that treatment with putrescine can partially alleviate oxidative damage caused by gamma rays.