Pontibacter qinzhouensis sp. nov., isolated from rhizosphere soil of a mangrove plant Rhizophora stylosa.

A novel bacterium of the genus Pontibacter, designated GY10130T, was isolated from rhizosphere soil of a mangrove plant Rhizophora stylosa collected from Guangxi province, China. Strain GY10130T was Gram-stain negative, positive for oxidase activities, aerobic, short rod-shaped cells without flagella. Growth was observed at 10-40 °C (optimum, 28 °C), pH 6.0-9.0 (optimum, 7.0) and NaCl concentrations of 0-4% (optimum, 1%). Strain GY10130T is closely related to members of the genus Pontibacter, namely P. beigongshangensis CGMCC 1.17104T (97.8%) and P. amylolyticus CGMCC 1.12749T (95.0%), P. humi SWU8T (94.7%), and less than 94.0% with other currently described type strains of Pontibacter. The strain GY10130T showed an ANI value of 80.6% and dDDH value of 23.2% with P. beigongshangensis CGMCC 1.17104T, followed by P. amylolyticus CGMCC 1.12749T with ANI and dDDH values of 72.9 and 13.8%, respectively. Strain GY10130T contains carotenoid-like pigments, but flexirubin-type pigments were absent. The cellular fatty acids (> 10%) consist of summed feature 4 (17:1 iso I/anteiso B) and iso-C15:0. The predominant menaquinone is MK-7. The polar lipids comprise phosphatidylethanolamine, two unidentified glycolipids, two unidentified aminolipids and six unidentified phospholipids. The genome length of strain GY10130T was 6.2 Mbp with a DNA G + C content of 47.1 mol% and 4727 protein-coding genes. The result of polyphasic taxonomic study show strain GY10130T represents one novel species of Pontibacter, Pontibacter qinzhouensis sp. nov., with the type strain GY10130T (=NBRC 113901T = CGMCC 1.16772T).

Unearthed New Indel in the Waxy Gene in Glutinous Rice Landraces from Thailand.

<b>Background and Objective:</b> Glutinous rice landraces have played important role in northeastern and northern region of Thailand and the Lao People's Democratic Republic (PDR) by ethnic groups in this area, both as staple food and ritual in rice culture since ancient times. At a present number of these rice, cultivars have decreased and disappeared from villages without considerations and/or explore DNA variation. This study compared target DNA sequences of <i>waxy</i> genes of a collection of glutinous rice landraces both upland and lowland rice. <b>Materials and Methods:</b> A collection of 50 glutinous rice landraces was explored DNA variation in the <i>Waxy</i> gene by re-sequencing DNA of the two segments (i.e., promoter, exon1 and intron1) of a gene. <b>Results:</b> New InDel of two deletion G at position 11 (GG-) and C at position 15 (CC-) were observed in DNA sequences of the promoter region and 5'untranslalted region of both upland and lowland rice accessions. Further, the (CT)₁₇ and (CT)₁₈ were two alleles in these glutinous rice landraces from northern and northeastern Thailand. All glutinous rice landraces exhibited a characteristic of low amylose content or glutinous rice: T Single Nucleotide Polymorphisms (SNP) (AGTTATA) in the first intron splice site. <b>Conclusion:</b> New InDel in DNA sequences of the promoter region of the <i>waxy</i> gene in glutinous rice landraces was first reported. This implies that it may reflect the status of genetic background in glutinous rice landraces in Thailand.

Pericarp color and haplotype diversity in weedy rice (O. sativa f. spontanea) from Thailand.

Weedy rice (Oryza sativa f. spontanea) is an important resource for breeding and evolutionary studies. The present study was carried out to identify the genotype of weedy rice strains responsible for variation of pericarp color by sequencing exon 6 of the Rc gene. Haplotype analysis revealed that rice strains with light red- and red-pericarp lacked the 14 bp deletion in the Rc gene, while rice strains with white-, brown and light green-pericarp contained the 14 bp deletion. A single nucleotide polymorphism (C-->A SNP) in exon 6 of the Rc gene was not found in all the weedy rice strains examined. This indicates that white pericarp allele in weedy rice gene pools of today might have originated by gene flow from locally grown modern rice varieties with white pericap into established gene pools of wild rice.

Molecular analysis of utility of a retrotransposon, p-SINE1-r2 in the Asian wild rice and weedy rice populations.

The distribution of a retrotransposon, p-SINE1-r2 located at the waxy locus was analyzed by the PCR assay in the perennial wild rice (Oryza rufipogon) which inhabited in four isolated and six disturbed populations and in the weedy rice population. The level of clonality of the wild rice species was determined in populations subject to level of water supply and another disturbance. The results showed that all four isolated populations carried the genotype (-/-) and (-/+), while three genotypes (-/-), (-/+) and (+/+) was found on the six populations which grown near by rice fields. This finding was strongly supported the idea that the original wild rice populations of O. rufipogon exhibited prominent genotype (-/-) and (-/+) and mainly propagated by vegetative reproduction and the allele (+) which found in the wild rice plant with the genotype (+/+) may originated from gene flow from cultivated rice to wild rice. Weedy rice accessions used in this study showed the three genotypes based on this DNA locus. The distribution of this DNA locus in wild rice and weedy rice populations were deviated from the Hardy-Weinberg equilibrium. The perennial wild rice populations were annually under season drought (March to May of the year in Thailand, Laos and Cambodia), they tended to have small size clones with relatively high clonal diversity (i.e., number of genotypes), except for the population from Cambodia, which carried only the genotype (-/+). Although DNA maker used to detect genetic variation at population levels is too small, but this locus is very sensitive enough to be a useful indicator for genetic variation at the population level.

Genetic variation of wild rice populations from Thailand and the Lao PDR based on molecular markers.

The two wild rice species, Oryza rufipogon and O. nivara, which inhabit Thailand and the Lao PDR, are threatened with the loss of their natural habitats. The losses are primarily attributable to human intervention. Before this process advances, it is crucial to obtain basic information on the genetic variations of these species, so important knowledge about the current status of genetic variability. In this study, genetic variation within and between fourteen natural populations of the two populations from Thailand and Lao PDR was investigated at the DNA level by analysis of Polymerase Chain Reaction (PCR) mediate molecular markers. The results illustrated that wild rice from Thailand carried deletion (D) type ORF 100, but deletion and non-deletion type (ND) were found in wild rice samples from the Lao PDR. Five different plastid subtypes (7C7A, 6C8A, 7C6A, 6C7A and 9C7A) were found in the collected samples. Both polymorphism and pattern of distribution of p-SINE1-r2 in the two wild rice species were found among the populations. These data indicated that genetic variation existed in these natural populations of wild rice, suggesting that the strategies should be developed that are conducive to the conservation of wild rice in its natural environment.

Identification of variant transcripts of waxy gene in non-glutinous rice (O. sativa L.) with different amylose content.

The identification of a number of cDNA sequences at the splice sites of Waxy (Wx) gene in rice endosperm point to the variation of gene expression associated with the G to T mutation at the 5' splice site of intron 1 and the alternative splicing produces variants transcripts by shortening. As consequence, variation of amylose content in this tissue. To determine whether the splicing type of Wx genes are also found in rice endosperm of Thai rice cultivars, cDNA of the gene have been isolated by using RT-PCR and sequenced. Analysis of the twenty-five cDNA clones led to the identification of the splicing pattern of the gene, four cryptic splice sites was found in cDNA sequences of low-amylose cultivars (<20%), while two cryptic splice sites of intermediate (20-25%) and high (>25%) amylose cultivars were characterized. The alternatively spliced transcripts were observed in much higher proportion in low amylose cultivars than that in intermediate and high amylose cultivars. These findings raise a possibility explanation that the degree of amylose content in non-glutinous rice cultivars may reflect to the amount of efficiently spliced transcripts.

An assessment of Wx microsatellite allele, alkali degradation and differentiation of chloroplast DNA in traditional black rice (Oryza sativa L.) from Thailand and Lao PDR.

Thailand and Lao PDR are the country's rich rice diversity. To contribute a significant knowledge for development new rice varieties, a collection of 142 black rice (Oryza sativa) accessions were determined for variation of physico-chemical properties, Wx microsatellite allele, Wx allele and chloroplast DNA type. The results showed that amylose content of black rice accessions were ranged from 1.9 to 6.8%. All of the alkali disintegration types (high, intermediate and low) was observed in these rice with average of 1.75 on the 1-3 digestibility scale. The unique Wx microsatellite allele (CT)17 was found in these samples and all black rice strains carried Wx(b) allele. In addition, all black rice accessions were found the duplication of the 23 bp sequence motif in the exon 2 of the wx gene. This evidence is a common phenomenon in glutinous rice. Based on two growing condition for black rice, rainfed lowland and rainfed upland, chloroplast DNA type was distinct from each other. All rice strains from rainfed lowland was deletion plastotype, but all other rainfed upland strains were non-deletion types.