Accumulation of Health-Promoting Compounds in Upland Black Rice by Interacting Mycorrhizal and Endophytic Fungi.

There is an increasing interest in finding eco-friendly and safe approaches to increase agricultural productivity and deliver healthy foods. Arbuscular mycorrhizal fungi (AMF) and endophytic fungi (EPF) are important components of sustainable agriculture in view of their ability to increase productivity and various plant secondary metabolites with health-promoting effects. In a pot experiment, our main research question was to evaluate the additive and synergistic effects of an AMF and four root-endophytic fungi on plant performance and on the accumulation of health-promoting secondary compounds. Plant growth varied between the treatments with both single inoculants and co-inoculation of an AMF and four EPF strains. We found that inoculation with a single EPF positively affected the growth and biomass production of most of the plant-endophyte consortia examined. The introduction of AMF into this experiment (dual inoculation) had a beneficial effect on plant growth and yield. AMF, Rhizophagus variabilis KS-02 co-inoculated with EPF, Trichoderma zelobreve PBMP16 increased the highest biomass, exceeding the growth rate of non-inoculated plants. Co-inoculated R. variabilis KS-02 and T. zelobreve PBMP16 had significantly greater beneficial effects on almost all aspects of plant growth, photosynthesis-related parameters, and yield. It also promoted root growth quality and plant nutrient uptake. The phenolic compounds, anthocyanin, and antioxidant capacity in rice seeds harvested from plants co-inoculated with AMF and EPF were dramatically increased compared with those from non-inoculated plants. In conclusion, our results indicated that EPF and AMF contributed to symbiosis in Maled Phai cultivar and were coordinately involved in promoting plant growth performance under a pot trial.

The Potential of Endophytic Fungi for Enhancing the Growth and Accumulation of Phenolic Compounds and Anthocyanin in Maled Phai Rice (Oryza sativa L.).

Rice (Oryza sativa L.) is one of the most popular cereal crops, being consumed by almost half of the world's population. Among several cultivars widely distributed in Thailand, Maled Phai is a Thai pigmented-upland rice with exceptionally high nutritional value and high demand in the local Thai market. This study aimed to investigate the feasibility of producing plant growth-promoting properties (PGP) and enhancing the accumulation of phytochemicals in Maled Phai rice seeds of endophytic fungi isolated from upland black rice. Among a total of 56 isolates, the 4 most effective PGP isolates were identified as Trichoderma zelobreve PBMP16, Talaromyces pinophilus PBMP28, Aspergillus flavus KKMP34, and Trichoderma sp. PBMP4 based on their morphological characteristics and multigene (ITS, rpb2, tef-1, CaM, and BenA) phylogenetic analyses. These four endophytic fungi could promote plant growth parameters under greenhouse conditions. Outstandingly, upland rice inoculated with Tr. zelobreve PBMP16 had a significant increase in total seed weight, root length, phenolic compounds, anthocyanin, antioxidants, and N uptake, which were higher than those of the noninoculated control, and even better than the chemical fertilizer. Overall, this report shows that endophytic fungi efficiently promoted growth and increased the phenolic compounds, anthocyanin, and antioxidants of Maled Phai rice.

High Temperature Alters Phenology, Seed Development and Yield in Three Rice Varieties.

Rice is an important and main staple food crop. Rice in Thailand grows in both the on- and off-seasons. The problem of growing rice in the off-season is that it is dry and the temperature tends to be high. To evaluate the effects of high temperatures on their phenology, yield and seed quality, three rice varieties were cultivated off-season in 2018 and 2019. Rice plants were grown in cement pots on planting date I (PDI; off-season; mid-January) and planting date II (PDII; late off-season; beginning of February). The results showed that rice plants were exposed to higher temperatures in 2019 (than 2018), as indicated by a higher accumulated growing degree day (AGDD). The high AGDD affected the phenology of the rice by shortening the duration of its development from sowing to physiological maturity (PM) from 106.8 DAS in 2018 to 86.0 DAS in 2019. The high AGDD shortened the development duration of the embryo and endosperm, resulting in reductions in the size and growth rates of the embryo and endosperm, and eventually reduced the yield and the yield components. Moreover, the high AGDD reduced the seed quality, as indicated by a decline in the seedling growth rate (SGR) and an increase in chalkiness. Among the varieties, the high temperature in 2019 caused the smallest phenological shift in Chai Nat 1 (CN1), while the shift was largest in Pathum Thani 1 (PTT1). In addition, CN1 exhibited a significantly higher total seed weight/panicle, 1000-seed weight and percentage of filled seed/pot than SP1 and PPT1. It was suggested that CN1 could be described as heat tolerant, and PTT1 as heat sensitive. It was also suggested that farmers should select appropriate rice varieties to grow in the off-season due to the risk of a high-temperature-induced reduction in the seed yield and quality.

The First Genetic Linkage Map of Winged Bean [Psophocarpus tetragonolobus (L.) DC.] and QTL Mapping for Flower-, Pod-, and Seed-Related Traits.

Winged bean [Psophocarpus tetragonolobus (L.) DC.] (2n = 2× = 18) is a tropical legume crop with multipurpose usages. Recently, the winged bean has regained attention from scientists as a food protein source. Currently, there is no breeding program for winged bean cultivars. All winged bean cultivars are landraces or selections from landraces. Molecular markers and genetic linkage maps are pre-requisites for molecular plant breeding. The aim of this study was to develop a high-density linkage map and identify quantitative trait loci (QTLs) for pod and seed-related traits of the winged bean. An F2 population of 86 plants was developed from a cross between winged bean accessions W054 and TPT9 showing contrasting pod length, and pod, flower and seed colors. A genetic linkage map of 1384 single nucleotide polymorphism (SNP) markers generated from restriction site-associated DNA sequencing was constructed. The map resolved nine haploid chromosomes of the winged bean and spanned the cumulative length of 4552.8 cM with the number of SNPs per linkage ranging from 36 to 218 with an average of 153.78. QTL analysis in the F2 population revealed 31 QTLs controlling pod length, pod color, pod anthocyanin content, flower color, and seed color. The number of QTLs per trait varied between 1 (seed length) to 7 (banner color). Interestingly, the major QTLs for pod color, anthocyanin content, and calyx color, and for seed color and flower wing color were located at the same position. The high-density linkage map QTLs reported in this study will be useful for molecular breeding of winged beans.

Dual-Purpose of the Winged Bean (Psophocarpus tetragonolobus (L.) DC.), the Neglected Tropical Legume, Based on Pod and Tuber Yields.

Winged beans (Psophocarpus tetragonolobus (L.) DC.) are grown as a vegetable legume crop in Thailand. All parts of the plant, including pods, seeds, leaves, flowers, and tubers are edible and are rich in protein and nutrients. Although the major consumption of winged bean is based on pod and tuber yields, only the people of Myanmar and Indonesia utilize winged bean tubers as food materials. The usefulness of the winged bean as an alternative crop for staple food and feed can shed some light on the impact of winged bean. Therefore, the evaluation of the dual purpose of the winged bean based on pod tuber yields is the objective of this study. In this study, ten-winged bean accessions-six accessions obtained from introduced sources and four accessions obtained from local Thai varieties-were laid out in randomized complete block design (RCBD) with three replications at the Agronomy Field Crop Station, Faculty of Agriculture, Khon Kaen University, Khon Kaen, Thailand from September 2019 to April 2020 and from October 2020 to April 2021. Data, including total pod weight, number of pods, pod length, 10-pod weight, and tuber weight were recorded, and the proximate nutrient and mineral contents in the tubers were also determined. The results revealed that the principal effects of year (Y) and genotype (G) were significant for total pod weight and the number of pods. Moreover, the Y × G interactions were principal effects upon the total pod weights and tuber weights. The results indicated that superior genotype and appropriate environmental conditions are key elements in successful winged bean production for both pod and tuber yields. The winged bean accessions W099 and W018 were consistent in both experimental years for pod and tuber yields at 23.6 and 18.36 T/ha and 15.20 and 15.5 T/ha, respectively. Each accession also proved high in tuber protein content at 20.92% and 21.04%, respectively, as well as significant in fiber, energy, and minerals. The results suggest that the winged bean accessions W099 and W018 can be used for dual-purpose winged bean production in Thailand.

Physiological and biochemical parameters for evaluation and clustering of rice cultivars differing in salt tolerance at seedling stage.

Salinity tolerance levels and physiological changes were evaluated for twelve rice cultivars, including four white rice and eight black glutinous rice cultivars, during their seedling stage in response to salinity stress at 100 mM NaCl. All the rice cultivars evaluated showed an apparent decrease in growth characteristics and chlorophyll accumulation under salinity stress. By contrast an increase in proline, hydrogen peroxide, peroxidase (POX) activity and anthocyanins were observed for all cultivars. The K(+)/Na(+) ratios evaluated for all rice cultivars were noted to be highly correlated with the salinity scores thus indicating that the K(+)/Na(+) ratio serves as a reliable indicator of salt stress tolerance in rice. Principal component analysis (PCA) based on physiological salt tolerance indexes could clearly distinguish rice cultivars into 4 salt tolerance clusters. Noteworthy, in comparison to the salt-sensitive ones, rice cultivars that possessed higher degrees of salt tolerance displayed more enhanced activity of catalase (CAT), a smaller increase in anthocyanin, hydrogen peroxide and proline content but a smaller drop in the K(+)/Na(+) ratio and chlorophyll accumulation.

A modified ferrous oxidation-xylenol orange assay for lipoxygenase activity in rice grains.

Ferrous oxidation-xylenol orange assay reagent was reformulated by using spectral analysis of ferric-xylenol orange complex to detect low concentrations of lipoxygenase rice grain products. Reducing the levels of ferrous sulphate and xylenol orange in the FOX reagent enabled the detection of low concentrations of hydroperoxy fatty acid derived from lipoxygenase activity in the range of 0.1-1.5 µM. Protein, substrate and time courses of the modified FOX assay were studied to determine lipoxygenase activity in rice grain. The assay was also applicable as a high throughput technique for comparisons of lipoxygenase activity from various rice varieties. This has important implications for rapid screening for low-lipoxygenase containing rice cultivars in rice breeding program and grain quality during storage.

Genetic diversity analysis of rice cultivars (Oryza sativa L. ) differing in salinity tolerance based on RAPD and SSR markers

Thirty rice cultivars were evaluated for salinity tolerance during the seedling stage and were divided into five tolerance groups including tolerant (T), moderately tolerant (MT), moderately susceptible (MS), susceptible (S) and highly susceptible (HS) which comprised 5, 10, 9, 4 and 2 cultivars respectively. Genetic diversity of all rice cultivars was evaluated using random amplified polymorphic DNA (RAPD) and simple sequence repeats (SSR) markers. The cultivars were evaluated for polymorphisms after amplification with 20 random decamer primers and 20 SSR primer pairs. A total of 161 RAPD markers and 190 SSR alleles were produced which revealed 68.94 percent and 89.47 percent polymorphism respectively. Mean genetic similarity coefficient was 0.82 for RAPD and 0.70 for SSR. Cluster analysis based on RAPD markers was effective in grouping cultivars based on their salt tolerance ability. Group IA1, IB and IV contained three T, three S and two HS rice cultivars respectively. The MT and MS cultivars which showed similar physiological responses to salinity were resolved into two groups: Group IA2 and Group II comprising ten and eight MT/MS cultivars respectively. Cluster analysis based on SSR markers separated rice cultivars into groups based on genetic relatedness which did not correspond to salinity tolerance level. The results from this study provided some useful implications for salt tolerance breeding programs. The evaluation of genetic similarity and cluster analysis together with salt tolerance ability provides some useful guides for assisting plant breeders in selecting suitable genetically diverse parents for the crossing program.