Early flowering, good grain quality mutants through gamma rays and EMS for enhancing per day productivity in rice (Oryza sativa L.).

PURPOSE: Rice is the prominent crop of world and it frames the important component of food chain. But, long duration of highly preferred rice varieties requires more water and are prone to biotic and abiotic stresses. Short duration rice varieties serve as a promising alternative in such cases. Hence, the present study was taken up to induce early flowering mutants in popular rice variety Bapatla 2231 (BPT 2231), a long duration medium slender rice variety using gamma rays and Ethyl Methane Sulfonate (EMS) treatment combinations. MATERIALS AND METHODS: Five hundred well filled and viable grains of optimum moisture content (12%) were subjected to irradiation with 8 doses comprising 100 Gy, 200 Gy, 300 Gy, 350 Gy, 400 Gy, 450 Gy, 500 Gy and 600 Gy treatments. For combination treatments, the irradiated seeds of gamma rays were further treated with 30 mM concentrations of Ethyl Methane sulfonate (EMS). The irradiated population were raised up to M3 generation for the identification of early maturing mutants. The isolated early maturing mutants in M3 generation were characterized for phenotypic, biochemical and grain qualities. RESULTS: The frequency of desirable early maturing mutants was observed more in combination treatments than the individual treatments of gamma rays. A total of 34 early maturing mutants (M) with variation for grain quality and biochemical traits were obtained. The mutants M-1, M-3, M-5, M-32, and M-34 were high yielding with fine grain type. The mutants M-4, M-17, M-18, M-19, M-20, and M-26 were high yielding with bold grain type. The mutants M-22, M-23, M-24, M-27, and M-28 were high in nutrient content. The early mutants matured 40-50 days earlier than control. CONCLUSION: In this variety, the combination treatments showed higher mutation frequency, mutagenic effectiveness and efficiency than the gamma rays in induction of both chlorophyll and viable mutants. This study revealed that the combination treatments were more effective in inducing early mutants than the gamma ray treatments alone. The early maturing mutants can be released as variety after evaluation under different yield trials and the mutants with increased nutrient content and varied grain quality can be utilized in hybridization/heterosis breeding programs for rice crop improvement.

Diverse role of phytic acid in plants and approaches to develop low-phytate grains to enhance bioavailability of micronutrients.

Natural or synthetic compounds that interfere with the bioavailability of nutrients are called antinutrients. Phytic acid (PA) is one of the major antinutrients present in the grains and acts as a chelator of micronutrients. The presence of six reactive phosphate groups in PA hinders the absorption of micronutrients in the gut of non-ruminants. Consumption of PA-rich diet leads to deficiency of minerals such as iron and zinc among human population. On the contrary, PA is a natural antioxidant, and PA-derived molecules function in various signal transduction pathways. Therefore, optimal concentration of PA needs to be maintained in plants to avoid adverse pleiotropic effects, as well as to ensure micronutrient bioavailability in the diets. Given this, the chapter enumerates the structure, biosynthesis, and accumulation of PA in food grains followed by their roles in growth, development, and stress responses. Further, the chapter elaborates on the antinutritional properties of PA and explains the conventional breeding and transgene-based approaches deployed to develop low-PA varieties. Studies have shown that conventional breeding methods could develop low-PA lines; however, the pleiotropic effects of these methods viz. reduced yield, embryo abnormalities, and poor seed quality hinder the use of breeding strategies. Overexpression of phytase in the endosperm and RNAi-mediated silencing of genes involved in myo-inositol biosynthesis overcome these constraints. Next-generation genome editing approaches, including CRISPR-Cas9 enable the manipulation of more than one gene involved in PA biosynthesis pathway through multiplex editing, and scope exists to deploy such tools in developing varieties with optimal PA levels.

Environmental impact of phytic acid in Maize (Zea mays. L) genotypes for the identification of stable inbreds for low phytic acid.

Phytic acid is a ubiquitous compound that chelates the micronutrients in food and hinder their absorption. Hence, breeding for low phytate content for producing stable low phytic acid (lpa) hybrids is essential. Phytic acid content in maize grains has been found to vary across environments and its stable expression has yet to be explored. In a view of this, forty inbreds were screened with two checks viz., CO-6 and CO-H(M)-8 across three locations. Twenty morphological and three quality traits were observed to identify the stable lines for low phytic acid with higher free inorganic phosphorous and starch. Among all the lines, UMI-467, LPA-2-285, LPA-2-395 and UMI-447 recorded a stable performance in both AMMI and GGE biplot analysis for low phytic acid (2.52-3.32 mg/g). These lines also had a higher free inorganic phosphorous, ensuring its bioavailability (1.78-1.88 mg/g). There were perturbations in yield, starch and seed characteristics of the stable low phytic acid lines due to their lower phytic acid concentrations. This stated the role of phytic acid in plant physiology and established the constraints to be faced in breeding for low phytic acid in maize. Among the lpa lines, LPA-2-285 (57.83%) and UMI-447 (55.78%) had the highest average starch content. The lowest stable phytic acid content was observed in UMI-467 (2.52 mg/g) and this line had severe reductions in yield parameters. Considering the seed and yield characteristics, LPA-2-285, LPA-2-395 and UMI-447 performed better than UMI-467. Although these four stable lines were poor in their adaptability among all the genotypes, they could be utilised as promising stable donors to facilitate the development of stable lpa hybrids.

Enumerating the phytic acid content in maize germplasm and formulation of reference set to enhance the breeding for low phytic acid.

Phytic acid (Myoinositol 1, 2, 3, 4, 5, 6 hexakisphosphate) is a ubiquitous compound present in plants. It is an important constituent in seed reducing the bioavailability of phosphorous and mineral nutrients when fed to monogastric animals like swine, poultry, fish etc. Hence, identification of maize germplasm with reduced phytic acid content is imperative to formulate the breeding programs to evolve low phytate lines. Towards this, three hundred and thirty-eight maize germplasm accessions available at Department of Millets, TNAU, were raised and screened for phytic acid content which varied from 2.77 to 16.70 mg/g of seed. Based on the variability present, a reference set with fifty-eight genotypes for phytic acid was formulated. The reference set was formed with random genotypes selected from the base population to follow a normal distribution (skewness; 0.17, kurtosis; 0.61 and K-S test for normality Dn = 0.70) for phytic acid. The non-significant difference between the means of the base and the reference ensured the entire representation of the base in the formulated reference for phytic acid. Among all the lines in the reference set, the lowest phytic acid content were observed in the lines UMI-113 (2.77 mg/g) followed by UMI-300-1 (3.17 mg/g), UMI-467 (5.50 mg/g) and UMI-158 (6.58 mg/g) could be used as donors for low phytic acid in breeding programs. The principal component analysis for studying the extent of variability in the reference, revealed six major principal components that exhibited 80.40% of variation with flowering traits, ear height and phytic acid as a major contributor for variability. The characters namely plant stand, germination percentage, kernel yield, ear length, ear diameter and number of kernels per row were found to be positively correlated with the phytic acid and this emphasizes the negative pleiotropic effects of low phytic acid lines in germination and seed set. Thus this formulated reference set enables the breeders to handle minimum population for further grouping the genotypes to analyse their heterotic potential combined with low phytic acid.

Gene interactions and genetics of blast resistance and yield attributes in rice (Oryza sativa L.).

Blast disease caused by the pathogen Pyricularia oryzae is a serious threat to rice production. Six generations viz., P1, P2, F1, F2, B1 and B2 of a cross between blast susceptible high-yielding rice cultivar ADT 43 and resistant near isogenic line (NIL) CT13432-3R, carrying four blast resistance genes Pi1, Pi2, Pi33 and Pi54 in combination were used to study the nature and magnitude of gene action for disease resistance and yield attributes. The epistatic interaction model was found adequate to explain the gene action in most of the traits. The interaction was complementary for number of productive tillers, economic yield, lesion number, infected leaf area and potential disease incidence but duplicate epistasis was observed for the remaining traits. Among the genotypes tested under epiphytotic conditions, gene pyramided lines were highly resistant to blast compared to individuals with single genes indicating that the nonallelic genes have a complementary effect when present together. The information on genetics of various contributing traits of resistance will further aid plant breeders in choosing appropriate breeding strategy for blast resistance and yield enhancement in rice.

A simple and rapid single kernel screening method to estimate amylose content in rice grains.

INTRODUCTION: In rice breeding programmes large number of grain samples are routinely analysed for amylose content (AC) through a tedious spectrophotometric method that also involves high reagent costs. OBJECTIVE: Here, we propose a rapid and economic screening technique for assessment of AC based on the amylose-iodine complex formation in the cut grains of rice, which we refer to as the cut grain dip (CGD) method. METHODS: The CGD method involves cutting the rice kernels in the middle with a pair of scissors and dipping the cut end in an optimised iodide:iodine (KI-I) solution termed the rapid amylose detection solution (RADS). RESULTS: It was found that the time taken for deep blue colouration by the cut end of the grains after dipping in RADS was proportional to the AC. The CGD method was further validated in a large set of rice mutants with varied AC. CONCLUSION: The proposed method can be used to screen samples for AC rapidly, with a single rice caryopsis, without any costly equipment and can be especially suitable for screening of mutants and segregants with altered AC in large breeding populations.

Epigenetic responses to drought stress in rice (Oryza sativa L.).

Cytosine methylation polymorphism plays a key role in gene regulation, mainly in expression of genes in crop plants. The differential expression of cytosine methylation over drought stress response was analyzed in rice using drought susceptible but agronomically superior lines IR 20 and CO 43, and drought tolerant genotypes PL and PMK 3 and their F1 hybrids. The parents and hybrids were subjected to two moisture regimes viz., one under drought condition and another under control condition. The cytosine methylation polymorphism in genomic DNA was quantified under both the conditions at the reproductive stage of the plant using the Methylation Sensitive Amplified Polymorphism (MSAP) technique devised by Xiong et al. (261:439-446, 1999). The results depicted that under drought condition, hyper-methylation was predominant in the drought susceptible genotypes while drought tolerant genotypes presented hypo-methylation behavior. While imposing drought, spikelet sterility per cent was positively correlated to percentage of methylation whereas, panicle length, number of seed per panicle, panicle weight, 100 seed weight, and yield/plant were negatively correlated indicating the role of epigenetic regulation in yield attributing traits in response to drought. Thus, methylation can be considered as an important epigenetic regulatory mechanism in rice plants to adapt drought situation. From this study, we speculate that the hyper- methylation may be an indicator of drought susceptibility and the hypo-methylation for drought tolerance and this methylation polymorphism can be effectively used in drought screening program.