Development of Gene-Based InDel Markers on Putative Drought Stress-Responsive Genes and Genetic Diversity of Durian (Durio zibethinus).

Insertion-deletion (InDel) markers are co-dominant, relatively abundant and practical for agarose gel genotyping. InDel polymorphism usually affects gene functions. Nucleotide sequences of durian (Durio zibethinus) are available, but InDel makers have not been well established. This study aimed to develop drought-related gene-based InDel markers for durian through bioinformatic analysis of RNA-Seq datasets. The polymorphism of the markers was verified in 24 durian genotypes local to Thailand. Bioinformatic analysis indicated 496 InDel loci having lengths more than 9 bp. To evaluate these InDel markers, 15 InDel loci were selected. Nine markers were successfully amplified a clear polymorphic band pattern on 2% agarose gel. The polymorphic information content (PIC) of these nine markers ranged from 0.1103 to 0.5808. The genetic distance between the 24 genotypes ranged from 0.222 to 0.889. The phylogeny based on the nine InDel loci distinguished the 24 genotypes and divided samples into four groups. This set of gene-based InDel markers on putative drought-responsive genes will be useful for genetic studies.

Responses of Lowland Rice Genotypes under Terminal Water Stress and Identification of Drought Tolerance to Stabilize Rice Productivity in Southern Thailand.

Lowland rice is an important cereal crop that plays a key role in the food security and the economy of Thailand. Terminal water stress (TWS) in rainfed lowland areas poses threats to rice productivity due to stress occurrence at terminal crop stages and extreme sensitivity of rice to TWS. A two-year study was conducted to characterize the performance of yield and yield attributes of twelve Thai lowland rice genotypes under TWS, to identify stress-tolerant genotypes using stress response indices and to identify promising stress indices which are correlated with grain yield (GY) under well-watered (WW) and TWS conditions for their use as rapid identifiers in a rice crop breeding program for enhancing drought stress tolerance. Measurements were recorded under WW and TWS conditions. Highly significant variations were observed amongst assessed genotypes for their yield productivity responses. According to stress response indices, genotypes were categorized into stress-tolerant and stress susceptible genotypes. Genotypes Hom Pathum, Sang Yod, Dum Ja and Pathum Thani-1 were found highly stress tolerant and relatively high yielding; genotypes Look Pla and Lep Nok were stress tolerant, whereas genotypes Chor Lung, Hom Nang Kaew and Hom Chan were moderately tolerant genotypes. Hence, stress-tolerant genotypes could be potentially used for cultivation under rainfed and water-limited conditions, where TWS is predicted particularly in southern Thailand to stabilize rice productivity. Stress tolerance indices, including stress tolerance index (STI), geometric mean productivity (GMP), mean productivity index (MPRO) and harmonic mean index (MHAR), indicated strong and positive associations with GY under WW and TWS; thus, these indices could be used to indicate stress tolerance in rice crop breeding program aimed at a rapid screening of lowland rice genotypes for stress tolerance.

Genotypic variation in 9-Cis-Epoxycarotenoid Dioxygenase3 gene expression and abscisic acid accumulation in relation to drought tolerance of Hevea brasiliensis.

Abscisic acid (ABA) is a stress-related plant hormone, which is reported to confer drought tolerance. A key enzyme in ABA biosynthesis is 9-cis-epoxycarotenoid dioxygenase. In this study, changes in morphological, physiological response, HbNCED3, and ABA accumulation of RRIM 623 and PB 5/51 rubber clones were observed at different time points of water deficit conditions (0, 3, 5, 7, and 9 days of withholding water). During water deficit, the relative water content (RWC), photosynthetic rate (Pn), and stomatal conductance (Gs) decreased, whereas the electro leakage (EL) increased. The magnitudes of the changes in these parameters were greater for PB 5/51 than for RRIM 623. Therefore, RRIM 623 was designated as representative of drought-tolerant clone and PB 5/51 as a drought-sensitive clone. The HbNCED3 transcription level of RRIM 623 showed lower expression compared with that of PB 5/51, which corresponded to the accumulation of ABA. RRIM 623 accumulated less ABA than PB 5/51. The ABA in RRIM 623 gradually increased, especially on the 7th day of withholding water, whereas that in PB 5/51 rapidly increased during the early periods of drought conditions. Additionally, the sensitivity of stomatal response to ABA showed that RRIM 623 had a higher sensitivity than PB 5/51. These results demonstrate that the drought-tolerant rubber clone, RRIM 623, was characterized by lower ABA accumulation during drought stress than the drought-sensitive clone, PB 5/51. The drought tolerance mechanism of the RRIM 623 might be associated with stomatal sensitivity to ABA accumulation under drought stress.

Overcoming Encouragement of Dragon Fruit Plant (Hylocereus undatus) against Stem Brown Spot Disease Caused by Neoscytalidium dimidiatum Using Bacillus subtilis Combined with Sodium Bicarbonate.

The use of the supernatant from a Bacillus subtilis culture mixed with sodium bicarbonate was explored as a means of controlling stem brown spot disease in dragon fruit plants. In in vitro experiments, the B. subtilis supernatant used with sodium bicarbonate showed a strong inhibition effect on the growth of the fungus, Neoscytalidium dimidiatum, the agent causing stem brown spot disease and was notably effective in preventing fungal invasion of dragon fruit plant. This combination not only directly suppressed the growth of N. dimidiatum, but also indirectly affected the development of the disease by eliciting the dragon-fruit plant's defense response. Substantial levels of the pathogenesis-related proteins, chitinase and glucanase, and the phenylpropanoid biosynthetic pathway enzymes, peroxidase and phenyl alanine ammonia-lyase, were triggered. Significant lignin deposition was also detected in treated cladodes of injured dragon fruit plants in in vivo experiments. In summary, B. subtilis supernatant combined with sodium bicarbonate protected dragon fruit plant loss through stem brown spot disease during plant development in the field through pathogenic fungal inhibition and the induction of defense response mechanisms.

Cloning and Expression Analysis of HbPR-1b and HbPR-3 in Hevea brasiliensis During Inoculation with Rigidoporus microporus.

BACKGROUND AND OBJECTIVE: Pathogenesis-related (PR) proteins are dramatically accumulated after pathogen infection. Innate defense response through increasing PR-proteins is important for rubber rootstock selection that is tolerant to the white root disease caused by Rigidoporus microporus. This study was aimed to investigate the expression levels of PR-1 and PR-3 genes in tolerant (PB5/51) and susceptible (BPM24 and RRIM600) rubber clones after R. microporus infection. MATERIALS AND METHODS: The mRNA of HbPR-1b and HbPR-3 was isolated and characterized from rubber leaves. Gene expression levels of HbPR-1b and HbPR-3 were compared among three rubber clones (PB5/51, BPM24 and RRIM600) after R. microporus infection at 0, 12, 24, 48, 72 and 96 h using quantitative real-time PCR. The relative transcript abundances between inoculated and control plants were compared using the means of gene expression between time points and by Tukey's HSD test. A probability value (p<0.05) was used to determine the significance of difference between time points. RESULTS: The open reading frame of HbPR-1b is 492 bp with deduced 163 amino acid residues and the phylogenetic analysis showed it shared significant evolutionary history and clustering into group I of PR-protein. Moreover, the partial HbPR-3 was isolated with 390 bp. Gene expression levels of HbPR-1b and HbPR-3 showed marked differences in both transcripts depending on the rubber clones. Two genes demonstrated up-regulation of both tolerance and susceptibility in response to attack by R. microporus. The highest expression levels were found in seedlings of PB5/51 after inoculation. In RRIM600, low expression levels of HbPR-1b and HbPR-3 were initially observed but gradually increased at 24 h post inoculation. The transcription profile of HbPR-1b was stable expression in BPM24. CONCLUSION: The results demonstrated that the level ofHbPR-1b and HbPR-3 transcription can distinguish between tolerant and susceptible clones. The candidate defense genes to the white root disease were observed in PB5/51 seedlings, particularly HbPR-1b.

Facultative Apomixis in Garcinia atroviridis (Clusiaceae) and Effects of Different Pollination Regimes on Reproductive Success.

Various aspects of the reproductive success of Garcinia atroviridis Griff. were studied. Controlled pollination experiments were carried out in an orchard located in Songkhla province, southern Thailand, from February to July 2003. Floral longevity, stigma receptivity, and pollen viability were examined before carrying out the experiments. Three pollination treatments were compared: open pollination, manual pollination with bags, and bags without pollination (apogamy). Although there was no significant difference in the initial fruit set, bagged and manual pollination produced a significantly greater fruit drop rate than apogamy or natural pollination at one week after the flowers had been pollinated. On the other hand, the apogamy treatment had a greater fruit drop rate than natural and manual pollination treatments before fruit maturation. In addition, unpollinated bagged flowers bore fewer and smaller fruit than naturally and manually cross-pollinated flowers. Although the fruits from unpollinated flowers were capable of asexual seed formation, they produced fewer seeds and had poorer seed quality (defined as average fresh weight and germination rate) than those from the other treatments. The occurrence of asexual and sexual reproduction was also studied using Random Amplification of Polymorphic DNA (RAPD) analysis and by comparing the patterns of bands produced from DNA extracted from the offspring of the naturally cross-pollinated fruits. On average, 58% of the offspring had a genetic constitution identical to that of the maternal parent (ranging from 36% to 87%), indicating that some offspring were produced without prior fertilisation. However, the remainder showed polymorphism, demonstrating the occurrence of sexual reproduction. These findings indicate that facultative apomixis occurred in the study population. However, a residual sexuality was important for fruit production, fruit size, normal seed set and seed quality.