Soybean (Glycine max) is a major plant source of protein and oil and produces important secondary metabolites beneficial for human health. As a tool for gene function discovery and improvement of this important crop, a mutant population was generated using fast neutron irradiation. Visual screening of mutagenized seeds identified a mutant line, designated MO12, which produced brown seeds as opposed to the yellow seeds produced by the unmodified Williams 82 parental cultivar. Using forward genetic methods combined with comparative genome hybridization analysis, we were able to establish that deletion of the GmHGO1 gene is the genetic basis of the brown seeded phenotype exhibited by the MO12 mutant line. GmHGO1 encodes a homogentisate dioxygenase (HGO), which catalyzes the committed enzymatic step in homogentisate catabolism. This report describes to our knowledge the first functional characterization of a plant HGO gene, defects of which are linked to the human genetic disease alkaptonuria. We show that reduced homogentisate catabolism in a soybean HGO mutant is an effective strategy for enhancing the production of lipid-soluble antioxidants such as vitamin E, as well as tolerance to herbicides that target pathways associated with homogentisate metabolism. Furthermore, this work demonstrates the utility of fast neutron mutagenesis in identifying novel genes that contribute to soybean agronomic traits.
Fungal volatile organic compounds (VOCs) emitted by Trichoderma species interact with a plant host and display multifaceted mechanisms. In this study, we investigated the antifungal activity of VOCs emitted by Trichoderma asperelloides PSU-P1 against fungal pathogens, as well as the ability of VOCs to activate defense responses and to promote plant growth in Arabidopsis thaliana. The strain’s VOCs had remarkable antifungal activity against fungal pathogens, with an inhibition range of 15.92–84.95% in a volatile antifungal bioassay. The VOCs of T. asperelloides PSU-P1 promoted the plant growth of A. thaliana, thereby increasing the fresh weight, root length, and chlorophyll content in the VOC-treated A. thaliana relative to those of the control. High expression levels of the chitinase (CHI) and β-1,3-glucanase (GLU) genes were found in the VOC-treated A. thaliana by quantitative reverse transcription polymerase chain reaction (RT-PCR). The VOC-treated A. thaliana had higher defense-related enzyme (peroxidase (POD)) and cell wall-degrading enzyme (chitinase and β-1,3-glucanase) activity than in the control. The headspace VOCs produced by PSU-P1, trapped with solid phase microextraction, and tentatively identified by gas chromatography–mass spectrometry, included 2-methyl-1-butanol, 2-pentylfuran, acetic acid, and 6-pentyl-2H-pyran-2-one (6-PP). The results suggest that T. asperelloides PSU-P1 emits VOCs responsible for antifungal activity, for promoting plant growth, and for inducing defense responses in A. thaliana.
The gene conferring a "pandan-like" aroma of winter melon was identified. The sequence variation (804-bp deletion) found in the gene was used as the target for functional marker development. Winter melon (Benincasa hispida), a member of the Cucurbitaceae family, is a commonly consumed vegetable in Asian countries that is popular for its nutritional and medicinal value. A "pandan-like" aroma, which is economically important in crops including rice and soybean, is rarely found in most commercial varieties of winter melon, but is present in some landraces. This aroma is a value-added potential trait in breeding winter melon with a higher economic value. In this study, we confirmed that the aroma of winter melon is due to the potent volatile compound 2-acetyl-1-pyrroline (2AP) as previously identified in other plants. Based on an analysis of public transcriptome data, BhAMADH encoding an aminoaldehyde dehydrogenase (AMADH) was identified as a candidate gene conferring aroma of winter melon. A sequence comparison of BhAMADH between the aromatic and non-aromatic accessions revealed an 804-bp deletion encompassing exons 11-13 in the aromatic accession. The deletion caused several premature stop codons and could result in a truncated protein with a length of only 208 amino acids compared with 503 amino acids in the normal protein. A functional marker was successfully developed based on the 804-bp deletion and validated in 237 F progenies. A perfect association of the marker genotypes and aroma phenotypes indicates that BhAMADH is the major gene conferring the aroma. The recently developed functional marker could be efficiently used in breeding programs for the aroma trait in winter melon.
The rapid expansion of the agricultural fi eld has had negative impacts on soil ecological functions. Soil fungi play an important role in nutrient cycling and the maintenance of soil functions in terrestrial ecosystems. However, our understanding of soil fungal communities and the interaction among microbial taxa in plant protected areas is still limited. This study applied high-throughput sequencing in an investigation of fungal communities and their diversity within the plant protected areas of Songkhla zoo in Southern Thailand. The sampling sites fell into three zones: zone I (dry evergreen forest, mixed deciduous forest and agricultural fi eld); zone II (evergreen forest and mixed deciduous forest); and zone III (evergreen forest, mixed deciduous forest and rubber tree crop). Soil samples were collected from each zone and subjected to physicochemical tests. Total DNA was extracted from soil samples, and we analyzed the nuclear ribosomal internal transcribed spacer 1 (ITS1) region of the rDNA gene of the fungi by Illumina Miseq sequencing to assess the fungal communities. The soil parameters revealed that the total calcium (Ca), manganese (Mg) and potassium (K) in zone III were signifi cantly higher than in the other zones. A total of 637, 702 and 745 OTUs were observed from zone I, zone II and zone III samples, respectively, with 143 OTUs common across all sampling sites. The fungal communities were signifi cantly different between the sampling sites. Zone III had the highest levels of community richness and diversity, followed by zone II and zone I. The dominant genera in zones I, II and III were Beauveria, Staphylotrichum and Trichoderma, respectively. Furthermore, the high concentration of K, Ca and Mg affected the fungal communities and diversity in the plant protected areas in Songkhla zoo. Our fi ndings provide insights into the importance of soil fungi, and this study identifi ed key differences in the fungal communities in the plant protected areas in Songkhla zoo.
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