Project description:The total RNA were extracted from pooled tissues of leaves and flowers from several plants of sacred lotus using TRIzol reagent (Invitrogen) according to the manufacturer's instructions. Then small RNAs ranging in 18–30 nucleotides were size fractionated electrophoretically, isolated from the gel, ligated with the 5′ and 3′ RNA adapters. The ligated product was reverse transcribed and subsequently amplified using 10–12 PCR cycles. The purified PCR product was sequenced using Illumina Genome Analyzer II. The qualified reads were used to predict phased small interfering RNAs from Chinese sacred lotus (Nelumbo nucifera Gaertn.).
Project description:Bacillus spp. and related genera native to soils of the pristine sacred groves from Meghalaya, India were characterized using biochemical and 16S rRNA gene analysis which revealed dominance of Bacillus, Paenibacillus, Lysinibacillus and Viridibacillus in the groves. Biochemical estimation was carried out for in vitro testing of plant growth promoting traits present in these isolates. PCR screening were performed for plant growth-promoting related genes involved in the biosynthesis of acid phosphatase (AcPho), indolepyruvate decarboxylase (ipdC), 1-aminocyclopropane-1-carboxylate deaminase (accd) and siderophore biosynthesis protein (asbA). 76% of the sacred grove isolates gave an amplified fragment for AcPho. Three of the isolates gave an amplified fragment for IpdC gene. Apart from 2 isolates, all the other isolates including the reference strains were positive for the amplification of the accd gene indicating their potential to produce ACC deaminase enzyme. 42% of the isolates gave an amplified fragment for asbA gene indicating the potential ability of these isolates to produce the catechol type siderophore, petrobactin. Overall findings indicated multiple PGP genetic traits present in these isolates which suggested that these isolates are capable of expressing multiple PGP traits. Phylogenetic and sequence analysis of accd and asbA genes from the isolates revealed that asbA genes from Paenibacillus taichungiensis SG3 and Paenibacillus tylopili SG24 indicated the occurrence of intergeneric horizontal transfer between Paenibacillus and Bacillus.
Project description:This study presents the complete genome sequence of Streptomyces californicus TBG-201 isolated from the soil samples of Vandanam sacred groves in Alleppey District, Kerala, India. The organism has potent chitinolytic activity. The genome of S. californicus TBG-201 was sequenced using the Illumina HiSeq-2500 platform with 2 × 150bp pair-end protocol and assembled using Velvet version 1.2.10.0. The assembled genome has a 7.99 Mb total length, a G+C content of 72.60%, and 6683 protein-coding genes, 116 pseudogenes, 31 rRNAs, and 66 tRNAs. AntiSMASH analysis revealed abundant biosynthetic gene clusters, while the dbCAN meta server was used to detect carbohydrate-active enzyme coding genes. The NCBI Prokaryotic Genome Annotation Pipeline was used for genome annotation. The presence of numerous genes coding for chitin degradation indicates the chitinolytic ability of this strain. The genome data have been deposited in NCBI with the accession number JAJDST000000000.
Project description:Very little is known about the mechanism controlling petiole rigidity in sacred lotus (Nelumbo nucifera Gaertn.). To investigate the mechanism controlling the lotus petiole rigidity, morphological and proteomic analyses were performed. Anatomically, there is a great variation between the petioles of floating and vertical leaves. The number of vascular bundles, ligneous cells and thickness of cell wall were higher in the initial vertical leaf petiole (IVP) compared to the initial floating leaf petiole (IFP). A total of 4855 proteins were quantified through comparative proteomic analysis, among which 421 proteins expressed 1.5 folds higher in IFP and 483 proteins expressed 1.5 folds higher in IVP. Protein function categories indicated hundreds of proteins involved in cell wall organization and biosynthesis, and cell wall assembly. Functional enrichment analysis for the differentially abundant proteins indicated the enrichment of 105 proteins in 6 different pathways, while 43 out the total proteins were enriched in lignin biosynthesis pathway. In consistent with genes and proteins expressions in lignin biosynthesis, the contents of lignin monomers precursors were significantly different in IFP and IVP. These findings support the involvement of lignin in lotus petioles rigidity.