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Fulltext Hereditary analysis and genotype × environment interaction effects on growth and yield components of Bambara groundnut (Vigna subterranea (L.) Verdc.) over multi-environments

Khan MMH, Rafii MY, Ramlee SI, Jusoh M, Al Mamun M

Sci Rep, 2022 09 19;12(1):15658.
PMID: 36123374 DOI: 10.1038/s41598-022-19003-z

This investigation was carried out to explore G × E interaction for yield and its associated attributes in 30 Bambara groundnut genotypes across four environments in tropical Malaysia. Such evaluations are essential when the breeding program's objective is to choose genotypes with broad adaption and yield potential. Studies of trait relationships, variance components, mean performance, and genetic linkage are needed by breeders when designing, evaluating, and developing selection criteria for improving desired characteristics in breeding programs. The evaluation of breeding lines of Bambara groundnut for high yield across a wide range of environments is important for long-term production and food security. Each site's experiment employed a randomized complete block design with three replicates. Data on vegetative and yield component attributes were recorded. The analysis of variance revealed that there were highly significant (p ≤ 0.01) differences among the 30 genotypes for all variables evaluated. A highly significant and positive correlation was identified between yield per hectare and dry seed weight (0.940), hundred seed weight (0.844), fresh pod weight (0.832), and total pod weight (0.750); the estimated correlation between dry weight of pods and seed yield was 1.0. The environment was more important than genotype and G × E in determining yield and yield components.A total of 49% variation is covered by PC1 (33.9%) and PC2 (15.1%) and the genotypes formed five distinct clusters based on Ward hierarchical clustering (WHC) method. The genotypes S5G1, S5G3, S5G5, S5G6, S5G8, S5G7, S5G2, S5G4, S5G10, S5G13, S5G11, and S5G14 of clusters I, II, and III were closest to the ideal genotype with superior yield across the environments. The PCA variable loadings revealed that an index based on dry pod weight, hundred seed weight, number of total pods and fresh pod weight could be used as a selection criteria to improve seed yield of Bambara groundnut.
Fulltext Path-coefficient and correlation analysis in Bambara groundnut (Vigna subterranea [L.] Verdc.) accessions over environments

Khan MMH, Rafii MY, Ramlee SI, Jusoh M, Al Mamun M

Sci Rep, 2022 01 07;12(1):245.
PMID: 34996953 DOI: 10.1038/s41598-021-03692-z

In a breeding program, studies of genotypic and phenotypic relationships among agricultural crop traits are useful to design, evaluate, and develop selection criteria for desirable traits. Using path coefficient analysis, the present study was executed to estimate the phenotypic, genotypic, and environmental correlation coefficients between yield and yield-related traits and to determine the direct and indirect effects of yield-related traits on yield per plant. A total of 30 genotypes of Vigna subterranea were studied under tropical conditions at two sites over two planting seasons (considered as four environments). The experiment at each site used a randomized complete block design with three replicates. Data were collected on vegetative and yield component attributes. Based on analysis of variance, pooled results showed that there were positive and highly significant differences (p ≤ 0.01) among the 30 genotypes for all attributes studied. Highly significant and positive strong correlation at phenotypic level was observed for dry seed weight (0.856), hundred seed weight (0.754), fresh pod weight (0.789), and total pod weight (0.626) with yield in kg per hectare, while moderate positive correlations were observed for harvest cut (0.360) and days to maturity (0.356). However, a perfect positive correlation was observed for the dry weight of pods with seed yield. In contrast, days to 50% flowering (- 0.350) showed a negative significant relationship with yield per hectare. The dried pod weight attribute (1.00) had a high positive direct effect on yield. Fresh pod weight had the greatest indirect effect on yield per hectare, followed by the number of total pods by dry pod weight. As a result, dry pod weight, hundred seed weight, number of total pods, and fresh pod weight could be used as selection criteria to improve the seed yield of Bambara groundnut (Vigna subterranea).
Fulltext AMMI and GGE biplot analysis for yield performance and stability assessment of selected Bambara groundnut (Vigna subterranea L. Verdc.) genotypes under the multi-environmental trails (METs)

Khan MMH, Rafii MY, Ramlee SI, Jusoh M, Al Mamun M

Sci Rep, 2021 Nov 23;11(1):22791.
PMID: 34815427 DOI: 10.1038/s41598-021-01411-2

The stability and high yielding of Vigna subterranea L. Verdc. genotype is an important factor for long-term development and food security. The effects of G × E interaction on yield stability in 30 Bambara groundnut genotypes in four different Malaysian environments were investigated in this research. The experiment used a randomized complete block design with three replications in each environment. Over multiple harvests, yield component traits such as the total number of pods per plant, fresh pods weight (g), hundred seeds weight (g), and yield per hectare were evaluated in the main and off-season in 2020 and 2021. Stability tests for multivariate stability parameters were performed based on analyses of variance. For all the traits, the pooled analysis of variance revealed highly significant (p
Molecular insight into genetic differentiation, population structure and banding pattern analysis of Bambara groundnut (Vigna subterranea [L.] Verdc.) linked with inter simple sequence repeats (ISSR)

Khan MMH, Rafii MY, Ramlee SI, Jusoh M, Al Mamun M, Kundu BC

Mol Biol Rep, 2023 Sep;50(9):7619-7637.
PMID: 37531035 DOI: 10.1007/s11033-023-08693-x

BACKGROUND: A set of 44 selected Bambara groundnut (Vigna subterranea [L.] Verdc.) accessions was sampled from 11 distinct populations of four geographical zones to assess the genetic drift, population structure, phylogenetic relationship, and genetic differentiation linked with ISSR primers.
METHODS AND RESULTS: The amplification of genomic DNA with 32 ISSR markers detected an average of 97.64% polymorphism while 35.15% and 51.08% polymorphism per population and geographical zone, respectively. Analysis of molecular variance revealed significant variation within population 75% and between population 25% whereas within region 84% and between region 16%. The Bidillali exposed greater number of locally common band i.e., NLCB (≤ 25%) = 25 and NLCB (≤ 50%) = 115 were shown by Cancaraki while the lowest was recorded as NLCB (≤ 25%) = 6 and NLCB (≤ 50%) = 72 for Roko and Maibergo, accordingly. The highest PhiPT value was noted between Roko and Katawa (0.405*) whereas Nei's genetic distance was maximum between Roko and Karu (0.124). Based on Nei's genetic distance, a radial phylogenetic tree was constructed that assembled the entire accessions into 3 major clusters for further confirmation unrooted NJ vs NNet split tree analysis based on uncorrected P distance exposed the similar result. Principal coordinate analysis showed variation as PC1 (15.04%) > PC2 (5.81%).
CONCLUSIONS: The current study leads to prompting the genetic improvement and future breeding program by maximum utilization and better conservation of existing accessions. The accessions under Cancaraki and Jatau are population documented for future breeding program due to their higher genetic divergence and homozygosity.
Fulltext DNA fingerprinting, fixation-index (Fst), and admixture mapping of selected Bambara groundnut (Vigna subterranea [L.] Verdc.) accessions using ISSR markers system

Khan MMH, Rafii MY, Ramlee SI, Jusoh M, Al Mamun M, Halidu J

Sci Rep, 2021 Jul 15;11(1):14527.
PMID: 34267249 DOI: 10.1038/s41598-021-93867-5

As a new crop in Malaysia, forty-four Bambara groundnut (Vigna subterranea L. verdc.) genotypes were sampled from eleven distinct populations of different origins to explore the genetic structure, genetic inconsistency, and fixation index. The Bambara groundnut, an African underutilized legume, has the capacity to boost food and nutrition security while simultaneously addressing environmental sustainability, food availability, and economic inequalities. A set of 32 ISSRs were screened out of 96 primers based on very sharp, clear, and reproducible bands which detected a total of 510 loci with an average of 97.64% polymorphism. The average calculated value of PIC = 0.243, RP = 5.30, H = 0.285, and MI = 0.675 representing the efficiency of primer set for genetic differentiation among the genotypes. The ISSR primers revealed the number of alleles (Na = 1.97), the effective number of alleles (Ne = 1.38), Nei's genetic diversity (h = 0.248), and a moderate level of gene flow (Nm = 2.26) across the genotypes studied. The estimated Shannon's information index (I = 0.395) indicates a high level of genetic variation exists among the accessions. Based on Nei's genetic dissimilarity a UPMGA phylogenetic tree was constructed and grouped the entire genotypes into 3 major clusters and 6 subclusters. PCA analysis revealed that first principal component extracted maximum variation (PC1 = 13.92%) than second principal component (PC2 = 12.59%). Bayesian model-based STRUCTURE analysis assembled the genotypes into 3 (best ΔK = 3) genetic groups. The fixation-index (Fst) analysis narrated a very great genetic diversity (Fst = 0.19 to 0.40) exists within the accessions of these 3 clusters. This investigation specifies the effectiveness of the ISSR primers system for the molecular portrayal of V. subterranea genotypes that could be used for genetic diversity valuation, detection, and tagging of potential genotypes with quick, precise, and authentic measures for this crop improvement through effective breeding schemes.
Fulltext Genetic analysis and selection of Bambara groundnut (Vigna subterranea [L.] Verdc.) landraces for high yield revealed by qualitative and quantitative traits

Khan MMH, Rafii MY, Ramlee SI, Jusoh M, Al Mamun M

Sci Rep, 2021 Apr 07;11(1):7597.
PMID: 33828137 DOI: 10.1038/s41598-021-87039-8

As a crop for the new millennium Bambara groundnut (Vigna subterranea [L.] Verdc.) considered as leading legumes in the tropical regions due to its versatile advantages. The main intent of this study was to find out the high yielding potential genotypes and considering these genotypes to develop pure lines for commercial cultivation in Malaysia. Considering the 14 qualitative and 27 quantitative traits of fifteen landraces the variation and genetic parameters namely, variability, heritability, genetic advance, characters association, and cluster matrix were determined. ANOVA revealed significant variation for all the agronomic traits (except plant height). Among the accessions, highly significant differences (P ≤ 0.01) were found for almost all the traits excluding fifty percent flowering date, seed length, seed width. The 16 traits out of the 27 quantitative traits had a coefficient of variation (CV) ≥ 20%. A positive and intermediate to perfect highly significant association (r = 0.23 to 1.00; P
Fulltext Genetic Diversity of Aromatic Rice Germplasm Revealed By SSR Markers

Jasim Aljumaili S, Rafii MY, Latif MA, Sakimin SZ, Arolu IW, Miah G

Biomed Res Int, 2018;2018:7658032.
PMID: 29736396 DOI: 10.1155/2018/7658032

Aromatic rice cultivars constitute a small but special group of rice and are considered the best in terms of quality and aroma. Aroma is one of the most significant quality traits of rice, and variety with aroma has a higher price in the market. This research was carried out to study the genetic diversity among the 50 aromatic rice accessions from three regions (Peninsular Malaysia, Sabah, and Sarawak) with 3 released varieties as a control using the 32 simple sequence repeat (SSR) markers. The objectives of this research were to quantify the genetic divergence of aromatic rice accessions using SSR markers and to identify the potential accessions for introgression into the existing rice breeding program. Genetic diversity index among the three populations such as Shannon information index (I) ranged from 0.25 in control to 0.98 in Sabah population. The mean numbers of effective alleles and Shannon's information index were 0.36 and 64.90%, respectively. Similarly, the allelic diversity was very high with mean expected heterozygosity (H
e
) of 0.60 and mean Nei's gene diversity index of 0.36. The dendrogram based on UPGMA and Nei's genetic distance classified the 53 rice accessions into 10 clusters. Analysis of molecular variance (AMOVA) revealed that 89% of the total variation observed in this germplasm came from within the populations, while 11% of the variation emanated among the populations. These results reflect the high genetic differentiation existing in this aromatic rice germplasm. Using all these criteria and indices, seven accessions (Acc9993, Acc6288, Acc6893, Acc7580, Acc6009, Acc9956, and Acc11816) from three populations have been identified and selected for further evaluation before introgression into the existing breeding program and for future aromatic rice varietal development.
Fulltext Genetic Diversity of Torch Ginger (Etlingera elatior) Germplasm Revealed by ISSR and SSR Markers

Ismail NA, Rafii MY, Mahmud TMM, Hanafi MM, Miah G

Biomed Res Int, 2019;2019:5904804.
PMID: 31198786 DOI: 10.1155/2019/5904804

Fifty-seven accessions of torch ginger (Etlingera elatior) collected from seven states in Peninsular Malaysia were evaluated for their molecular characteristics using ISSR and SSR markers to assess the pattern of genetic diversity and association among the characteristics. Diversity study through molecular characterization showed that high variability existed among the 57 torch ginger accessions. ISSR and SSR molecular markers revealed the presence of high genetic variability among the torch ginger accessions. The combination of different molecular markers offered reliable and convincing information about the genetic diversity of torch ginger germplasm. This study found that SSR marker was more informative compared to ISSR marker in determination of gene diversity, polymorphic information content (PIC), and heterozygosity in this population. SSR also revealed high ability in evaluating diversity levels, genetic structure, and relationships of torch ginger due to their codominance and rich allelic diversity. High level of genetic diversity discovered by SSR markers showed the effectiveness of this marker to detect the polymorphism in this germplasm collection.
Molecular markers: a potential resource for ginger genetic diversity studies

Ismail NA, Rafii MY, Mahmud TM, Hanafi MM, Miah G

Mol Biol Rep, 2016 Dec;43(12):1347-1358.
PMID: 27585572

Ginger is an economically important and valuable plant around the world. Ginger is used as a food, spice, condiment, medicine and ornament. There is available information on biochemical aspects of ginger, but few studies have been reported on its molecular aspects. The main objective of this review is to accumulate the available molecular marker information and its application in diverse ginger studies. This review article was prepared by combing material from published articles and our own research. Molecular markers allow the identification and characterization of plant genotypes through direct access to hereditary material. In crop species, molecular markers are applied in different aspects and are useful in breeding programs. In ginger, molecular markers are commonly used to identify genetic variation and classify the relatedness among varieties, accessions, and species. Consequently, it provides important input in determining resourceful management strategies for ginger improvement programs. Alternatively, a molecular marker could function as a harmonizing tool for documenting species. This review highlights the application of molecular markers (isozyme, RAPD, AFLP, SSR, ISSR and others such as RFLP, SCAR, NBS and SNP) in genetic diversity studies of ginger species. Some insights on the advantages of the markers are discussed. The detection of genetic variation among promising cultivars of ginger has significance for ginger improvement programs. This update of recent literature will help researchers and students select the appropriate molecular markers for ginger-related research.
Fulltext Genetic consequences of chronic gamma irradiation on agro morphological traits in chili under hydrogel enhance media

Hashim A, Rafii MY, Yusuff O, Harun AR, Juraimi S, Misran A, et al.

Heliyon, 2024 Feb 29;10(4):e25111.
PMID: 38370252 DOI: 10.1016/j.heliyon.2024.e25111

Induced mutation for the creation of desirable traits through chronic gamma irradiation provides an opportunity for the selection and development of new chili varieties. This study was conducted to assess the effects of different doses of chronic gamma irradiation on morpho-physiological traits in chili. Ten plants from each variety were exposed to different doses of chronic gamma irradiation for 277.02 h at three weeks after germination under gamma greenhouse facilities, with accumulative dose; 185.61Gy, 83.11Gy, 47.096Gy, 30.474Gy, 19.4Gy, 13.9Gy, 11.1Gy, 8.31Gy, 5.54Gy) and 2.77Gy respectively. Highly significant differences were observed among doses (Rings) of chronic gamma irradiation expressed in mean values for all investigated traits. Relatively moderate doses of chronic gamma irradiation represented by doses 47.096 Gy (Ring 4) and 19.40 Gy (Ring 6) resulted in significant stimulation for most of the studied characters. The highest heritability was recorded in days to flowering at 99.88 while the lowest was observed in fruit dry weight at 34.66 %. High genetic advance were recorded for most of the quantitative traits studied. In addition, a highly significant positive correlation was observed between total fruit per plant, total number of fruit per plant, plant height, fruit fresh weight, number of secondary branches, chlorophyll a, fruit dry weight, total chlorophyll content, stem diameter, fruit length and fruit girth. With increasing chronic gamma dose, mutagenic efficiency and efficacy generally increased. Induced variety of desirable features will considerably increase the chilli's amelioration through mutation breeding, leading to the development of improved varieties. The results of this research offer valuable information for the use of chronic gamma radiation in the mutations breeding of Capsicum annuum L., which will be advantageous for future breeding programs.
Genetic analysis and identification of SSR markers associated with rice blast disease in a BC2F1 backcross population

Hasan N, Rafii MY, Abdul Rahim H, Nusaibah SA, Mazlan N, Abdullah S

Genet. Mol. Res., 2017 Jan 23;16(1).
PMID: 28128411 DOI: 10.4238/gmr16019280

Rice (Oryza sativa L.) blast disease is one of the most destructive rice diseases in the world. The fungal pathogen, Magnaporthe oryzae, is the causal agent of rice blast disease. Development of resistant cultivars is the most preferred method to achieve sustainable rice production. However, the effectiveness of resistant cultivars is hindered by the genetic plasticity of the pathogen genome. Therefore, information on genetic resistance and virulence stability are vital to increase our understanding of the molecular basis of blast disease resistance. The present study set out to elucidate the resistance pattern and identify potential simple sequence repeat markers linked with rice blast disease. A backcross population (BC2F1), derived from crossing MR264 and Pongsu Seribu 2 (PS2), was developed using marker-assisted backcross breeding. Twelve microsatellite markers carrying the blast resistance gene clearly demonstrated a polymorphic pattern between both parental lines. Among these, two markers, RM206 and RM5961, located on chromosome 11 exhibited the expected 1:1 testcross ratio in the BC2F1 population. The 195 BC2F1 plants inoculated against M. oryzae pathotype P7.2 showed a significantly different distribution in the backcrossed generation and followed Mendelian segregation based on a single-gene model. This indicates that blast resistance in PS2 is governed by a single dominant gene, which is linked to RM206 and RM5961 on chromosome 11. The findings presented in this study could be useful for future blast resistance studies in rice breeding programs.
Marker-assisted backcrossing: a useful method for rice improvement

Hasan MM, Rafii MY, Ismail MR, Mahmood M, Rahim HA, Alam MA, et al.

Biotechnology, biotechnological equipment, 2015 Mar 04;29(2):237-254.
PMID: 26019637

The world's population is increasing very rapidly, reducing the cultivable land of rice, decreasing table water, emerging new diseases and pests, and the climate changes are major issues that must be addressed to researchers to develop sustainable crop varieties with resistance to biotic and abiotic stresses. However, recent scientific discoveries and advances particularly in genetics, genomics and crop physiology have opened up new opportunities to reduce the impact of these stresses which would have been difficult if not impossible as recently as the turn of the century. Marker assisted backcrossing (MABC) is one of the most promising approaches is the use of molecular markers to identify and select genes controlling resistance to those factors. Regarding this, MABC can contribute to develop resistant or high-yielding or quality rice varieties by incorporating a gene of interest into an elite variety which is already well adapted by the farmers. MABC is newly developed efficient tool by which using large population sizes (400 or more plants) for the backcross F1 generations, it is possible to recover the recurrent parent genotype using only two or three backcrosses. So far, many high yielding, biotic and abiotic stresses tolerance, quality and fragrance rice varieties have been developed in rice growing countries through MABC within the shortest timeframe. Nowadays, MABC is being used widely in plant breeding programmes to develop new variety/lines especially in rice. This paper reviews recent literature on some examples of variety/ line development using MABC strategy.
Introgression of blast resistance genes into the elite rice variety MR263 through marker-assisted backcrossing

Hasan MM, Rafii MY, Ismail MR, Mahmood M, Alam MA, Abdul Rahim H, et al.

J Sci Food Agric, 2016 Mar 15;96(4):1297-305.
PMID: 25892666 DOI: 10.1002/jsfa.7222

Blast caused by the fungus Magnaporthe oryzae is a significant disease threat to rice across the world and is especially prevalent in Malaysia. An elite, early-maturing, high-yielding Malaysian rice variety, MR263, is susceptible to blast and was used as the recurrent parent in this study. To improve MR263 disease resistance, the Pongsu Seribu 1 rice variety was used as donor of the blast resistance Pi-7(t), Pi-d(t)1 and Pir2-3(t) genes and qLN2 quantitative trait locus (QTL). The objective was to introgress these blast resistance genes into the background of MR263 using marker-assisted backcrossing with both foreground and background selection.
Novel SNPs in the SPAG11 gene and association with testicular biometric variables in Boer goats and application of the levelled-container technique

Harighi MF, Wahid H, Thomson PC, Rafii MY, Jesse FFA

Anim. Reprod. Sci., 2019 Sep;208:106113.
PMID: 31405472 DOI: 10.1016/j.anireprosci.2019.106113

Testicular volume (TV) is one of the most important traits used in evaluation of the reproductive capacity of male animals. The levelled-container used in the present study was found to be reliable instrument to measure TV, based on a water displacement method. Sperm-associated antigen 11 (SPAG11) is an important gene that affects male reproductive performance. An objective of the present study, therefore, was to determine if single nucleotide polymorphisms (SNPs) in a fragment of the SPAG11 gene could be used to determine associations with values of testicular biometric variables in Boer goats. Primers were designed to amplify the full length of the first two exons of SPAG11. The targeted fragment was generated using a molecular cloning technique. As the result, four SNPs, [g.1256A > G(ss19199134542), g.1270C > T(ss19199134541), g.1325A > G(ss19199134540) and g.1327 G > A (ss19199134543)], were detected using a single-base extension (SBE) method. Two of these SNPs were synonymous (ss19199134540 and ss19199134542). The other two SNPs were nonsynonymous, thus, there were changes in amino acid in the resulting protein: threonine to isoleucine (for ss19199134541) and arginine to glutamine (for ss19199134543). The SNP ss19199134543 was the only locus detected that was associated with TV (P = 0.002). None of the testes dimensions nor TW were associated with detected SPAG11 gene SNPs. Most likely, the ss19199134543 locus affects tissue structures adjacent to the testes, causing the change in TV. In conclusion, among the studied testicular biometric variables, TV had the greatest potential for preselecting of bucks with desirable semen quality. The use of the levelled-container as a TV measurement approach was an accurate and reliable method.
Flooding tolerance in Rice: adaptive mechanism and marker-assisted selection breeding approaches

Haque MA, Rafii MY, Yusoff MM, Ali NS, Yusuff O, Arolu F, et al.

Mol Biol Rep, 2023 Mar;50(3):2795-2812.
PMID: 36592290 DOI: 10.1007/s11033-022-07853-9

Natural and man-made ecosystems worldwide are subjected to flooding, which is a form of environmental stress. Genetic variability in the plant response to flooding involves variations in metabolism, architecture, and elongation development that are related with a low oxygen escape strategy and an opposing quiescence scheme that enables prolonged submergence endurance. Flooding is typically associated with a decrease in O2 in the cells, which is especially severe when photosynthesis is absent or limited, leading to significant annual yield losses globally. Over the past two decades, considerable advancements have been made in understanding of mechanisms of rice adaptation and tolerance to flooding/submergence. The mapping and identification of Sub1 QTL have led to the development of marker-assisted selection (MAS) breeding approach to improve flooding-tolerant rice varieties in submergence-prone ecosystems. The Sub1 incorporated in rice varieties showed tolerance during flash flood, but not during stagnant conditions. Hence, gene pyramiding techniques can be applied to combine/stack multiple resistant genes for developing flood-resilient rice varieties for different types of flooding stresses. This review contains an update on the latest advances in understanding the molecular mechanisms, metabolic adaptions, and genetic factors governing rice flooding tolerance. A better understanding of molecular genetics and adaptation mechanisms that enhance flood-tolerant varieties under different flooding regimes was also discussed.
Fulltext Physicochemical Properties and Tissue Structure of High Kernel Elongation Rice (Oryza sativa L.) Varieties as Affected by Heat Treatment

Halim AABA, Rafii MY, Osman MB, Chukwu SC, Oladosu Y

Foods, 2023 May 31;12(11).
PMID: 37297452 DOI: 10.3390/foods12112207

Heat treatment could affect the structure and properties of rice varieties. The present study was conducted in order to determine the effects of heat treatment on the physicochemical properties and tissue structure of Mahsuri Mutan, Basmati 370 and MR219 rice varieties. The three rice varieties were subjected to heat treatment (ageing) at 90 °C, using an oven, for 3 h. After the heat treatment, the samples were cooled at room temperature (25 °C) for 1 h. Physicochemical properties, such as alkali digestion value, water uptake ratio, solids in cooking water, high kernel elongation ratio and amylose contents, were determined. The procedure used in determining both apparent and absolute amylose involved measuring the iodine affinity of defatted whole starch. Ahigh-performance anion-exchange chromatograph was used to analyse branch chain length distribution of amylopectin quantitatively. The starch structure of the rice samples was observed under a scanning electron microscope. Data collected on physicochemical traits, heat treatment and control (ageing and non-ageing) were subjected to an analysis of variance using SAS software version 9.4. In this study, Mahsuri Mutan and Basmati 370 showed superior high kernel elongation as compared to their respective rice progenies. This study also found that heat treatment directly affected the increasingly high kernel elongation for both populations. The phenotypic correlation co-efficient indicated that there was a high positive correlation between high kernel elongation and water uptake ratio, implying that selection for water uptake ratio would increase the high kernel elongation characteristic. The heat treatment showed significant difference in all the physicochemical traits of the varieties studied. Heat treatment also affected the very long branch chains of starch, such as amylose. Observation under an electron microscope showed that the samples subjected to heat treatment had more cracks on the tissue structure compared to normal rice samples. The hexagon structure in Mahsuri Mutan produced a greater elongation effect on its kernel. The findings from this study could be useful to breeders in the selection and development of a new high kernel elongation rice variety.
The effect of salinity on chlorophyll, proline and mineral nutrients in common weeds of coastal rice fields in Malaysia

Hakim MA, Juraimi AS, Hanafi MM, Ismail MR, Rafii MY, Aslani F, et al.

J Environ Biol, 2014 Sep;35(5):855-64.
PMID: 25204059

Six weed species (Leptochola chinensis, Echinochloa crus-galli, Echinochloa colona, Jussiaea linifolia, Oryza sativa (weedy rice) and Cyperus iria) were tested for their salt tolerant traits in terms of chlorophyll, proline and mineral nutrients accumulation against different salinity levels (0, 4, 8, 16, 24, 32, and 40 dS m(-1)). Chlorophyll a, b and total chlorophyll content, proline and mineral nutrients accumulation were determined. Salt stress showed prominent effect on all the parameters investigated and there were significant variations between the all weed species. Chlorophyll content, K+, Ca(2+) and Mg(2+) ions in both shoots and roots significantly decreased; while proline and Na+ accumulation significantly increased with increasing salinity up to 40 dS m(-1). In terms of overall performance, Cyperus iria and E. crus-galliwere relatively more tolerant; E. colona and J. linifolia were tolerant; L. chinensis and O. sativa L were salt sensitive, respectively.
Fulltext Biochemical and anatomical changes and yield reduction in rice (Oryza sativa L.) under varied salinity regimes

Hakim MA, Juraimi AS, Hanafi MM, Ismail MR, Selamat A, Rafii MY, et al.

Biomed Res Int, 2014;2014:208584.
PMID: 24579076 DOI: 10.1155/2014/208584

Five Malaysian rice (Oryza sativa L.) varieties, MR33, MR52, MR211, MR219, and MR232, were tested in pot culture under different salinity regimes for biochemical response, physiological activity, and grain yield. Three different levels of salt stresses, namely, 4, 8, and 12 dS m(-1), were used in a randomized complete block design with four replications under glass house conditions. The results revealed that the chlorophyll content, proline, sugar content, soluble protein, free amino acid, and yield per plant of all the genotypes were influenced by different salinity levels. The chlorophyll content was observed to decrease with salinity level but the proline increased with salinity levels in all varieties. Reducing sugar and total sugar increased up to 8 dS m(-1) and decreased up to 12 dS m(-1). Nonreducing sugar decreased with increasing the salinity levels in all varieties. Soluble protein and free amino acid also decreased with increasing salinity levels. Cortical cells of MR211 and MR232 did not show cell collapse up to 8 dS m(-1) salinity levels compared to susceptible checks (IR20 and BRRI dhan29). Therefore, considering all parameters, MR211 and MR232 showed better salinity tolerance among the tested varieties. Both cluster and principal component analyses depict the similar results.
The genetic and molecular origin of natural variation for the fragrance trait in an elite Malaysian aromatic rice through quantitative trait loci mapping using SSR and gene-based markers

Golestan Hashemi FS, Rafii MY, Ismail MR, Mohamed MT, Rahim HA, Latif MA, et al.

Gene, 2015 Jan 25;555(2):101-7.
PMID: 25445269 DOI: 10.1016/j.gene.2014.10.048

MRQ74, a popular aromatic Malaysian landrace, allows for charging considerably higher prices than non-aromatic landraces. Thus, breeding this profitable trait has become a priority for Malaysian rice breeding. Despite many studies on aroma genetics, ambiguities considering its genetic basis remain. It has been observed that identifying quantitative trait loci (QTLs) based on anchor markers, particularly candidate genes controlling a trait of interest, can increase the power of QTL detection. Hence, this study aimed to locate QTLs that influence natural variations in rice scent using microsatellites and candidate gene-based sequence polymorphisms. For this purpose, an F2 mapping population including 189 individual plants was developed by MRQ74 crosses with 'MR84', a non-scented Malaysian accession. Additionally, qualitative and quantitative approaches were applied to obtain a phenotype data framework. Consequently, we identified two QTLs on chromosomes 4 and 8. These QTLs explained from 3.2% to 39.3% of the total fragrance phenotypic variance. In addition, we could resolve linkage group 8 by adding six gene-based primers in the interval harboring the most robust QTL. Hence, we could locate a putative fgr allele in the QTL found on chromosome 8 in the interval RM223-SCU015RM (1.63cM). The identified QTLs represent an important step toward recognition of the rice flavor genetic control mechanism. In addition, this identification will likely accelerate the progress of the use of molecular markers for gene isolation, gene-based cloning, and marker-assisted selection breeding programs aimed at improving rice cultivars.
Fulltext Application of an Effective Statistical Technique for an Accurate and Powerful Mining of Quantitative Trait Loci for Rice Aroma Trait

Golestan Hashemi FS, Rafii MY, Ismail MR, Mohamed MT, Rahim HA, Latif MA, et al.

PLoS One, 2015;10(6):e0129069.
PMID: 26061689 DOI: 10.1371/journal.pone.0129069

When a phenotype of interest is associated with an external/internal covariate, covariate inclusion in quantitative trait loci (QTL) analyses can diminish residual variation and subsequently enhance the ability of QTL detection. In the in vitro synthesis of 2-acetyl-1-pyrroline (2AP), the main fragrance compound in rice, the thermal processing during the Maillard-type reaction between proline and carbohydrate reduction produces a roasted, popcorn-like aroma. Hence, for the first time, we included the proline amino acid, an important precursor of 2AP, as a covariate in our QTL mapping analyses to precisely explore the genetic factors affecting natural variation for rice scent. Consequently, two QTLs were traced on chromosomes 4 and 8. They explained from 20% to 49% of the total aroma phenotypic variance. Additionally, by saturating the interval harboring the major QTL using gene-based primers, a putative allele of fgr (major genetic determinant of fragrance) was mapped in the QTL on the 8th chromosome in the interval RM223-SCU015RM (1.63 cM). These loci supported previous studies of different accessions. Such QTLs can be widely used by breeders in crop improvement programs and for further fine mapping. Moreover, no previous studies and findings were found on simultaneous assessment of the relationship among 2AP, proline and fragrance QTLs. Therefore, our findings can help further our understanding of the metabolomic and genetic basis of 2AP biosynthesis in aromatic rice.

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