Histological analysis, which aims to investigate the microscopic anatomy of biological tissues, has been a simple and powerful technique for plant taxonomy. Sectioning followed by staining methods is widely used in observing histological structures. However, the staining techniques often destroy tissue and provide low-quality images due to nonspecific reactions with the dyes making further analysis difficult. In this report, we propose an applicable non-staining histology protocol based on auto-fluorescence characteristics of plant tissues and its application in the anatomical discrimination of six similar-appearance species of golden camellias as a case study. We compared the images from the same tissue under a bright field with the staining step and under fluorescence directly without the staining step in the sample preparation. The images were taken from Eclipse Ni-U microscopy (Nikon, Japan) with a color DS-Ri2 camera (Nikon, Japan) and NIS-ELEMENTS Basic Research Imaging software. The non-staining method demonstrated significant advantages compared to the staining protocol. The fluorescent images showed the distinction between adjacent leaf tissues with their own naturally reflective colors. In addition, the anatomical parameters, including the xylem area, phloem area, bundle sheath area, and palisade/spongy width ratio, were easily measured in good-quality images. These parameters were used in discriminative analysis by the Principal Component Analysis (PCA). The PCA diagram demonstrated the separation of six species, thus suggesting that these anatomical parameters can be used for taxonomy. In conclusion, our study showed a helpful technique in histological analysis that significantly contributes to the taxonomy of golden camellias species and can be applied in other plant varieties.
Endophytic bacteria (EB) possess different beneficial traits. Endophytic microbes are often functional in that they may carry nutrients from the soil into plants, modulate plant development, increase stress tolerance of plants, suppress virulence in pathogens, increase disease resistance in plants, and suppress development of competitor plant species. They may enhance plant development by carrying nutrients from the soil into plants and protect plants against phytopathogens by synthesizing extra-/intracellular proteolytic enzymes, as well as releasing antimicrobial metabolites and competing with pathogens for habitation and nutrients. In this study, we investigated the ability to produce exo/endo-enzymes such as gelatinase, amylase, and catalase, and antagonistic activity of 77 EB strains isolated from lowland rice roots grown in Doan Ket commune, Thanh Mien district, Hai Duong province, Vietnam. Out of 77 isolates, 76 (98.71%) showed the ability to liquefy gelatin after 7 days with different rates of hydrolysis. The test of starch hydrolysis revealed 58 (75.33%) isolates that were able to hydrolyze starch. Fifty-one out of 77 isolates (66.24%) were able to produce catalase. The antagonistic activity of rice root endophytic bacteria was determined against bacterial leaf blight disease-causing pathogen Xanthomonas oryzae pv. oryzae (Xoo), strains X19.2 and VX41. We found that three isolates (TP5, TP7, TP11) showed the ability to inhibit the growth of strain VX41 and twelve isolates (TP3, TP7, TP8, TP9, TP10, TP11, TP12, TP13, TP15, TP17, TP21, TP23) were able to inhibit the growth of strain X19.2. These results are served as a venue for further investigation in planta under the conditions of net house and field trials in order to confirm the potential strains for the development of bioinoculant toward controlling the disease caused by Xoo.
Phosphorus is the one of the most important macro-elements for the growth, development as well as productivity of plants. However, the overuse of fertilizer negatively affects soil and water quality, and the run-out of natural phosphates (Pi). Therefore, it is necessary to study on Pi cycle and find out the way to use Pi efficiently for rice as well as other crops. In this work, genome wide association studies (GWAS) was used to investigate the diversity in uptaking phosphate of 157 Vietnamese rice cultivars in order to find genes involved in this process. Rice plants were grown in sand columns and irrigated with Yoshida nutrient medium every three days during 6 weeks. The experiment was conducted with three repetations in a randomized complete block design. The natural ability of uptaking the phosphate was then quantified and evaluated. GWAS analysis was conducted using the Mix Linear Model that combine both kinship and population architechture of the panel with 6 Principale Component as co-factors. Results obtained from the association mapping revealed a total of 19 important single nucleotide polymorphism (SNPs), 9 quantitative trait locus (QTLs) and 22 genes. The obtained findings may provide genetic tools for the development of rice varieties which can use phosphate effectively.
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