In this work chitin was extracted chemically from shrimp shells. Seventeen Bacillus isolates were screened for chitinolytic activity. The chitinolytic strains of Bt. were screened at different temperatures and pHs for their hydrolytic potentials. By using a pair of specific primers, endochitinase gene was amplified from SBS Bt-5 strain through PCR, and then cloned into pTZ57 TA cloning vector and transferred in Escherichia coli DH5α strain. The sequenced gene (GenBank Accession No: HE995800) consists of 2031 nucleotides capable of encoding 676 residues. The protein consisted of three functional domains with a calculated molecular mass of 74.53 kDa and a pI value of 5.83. The amino acid sequence of chi gene showed 99% similarity to the genes of Bt MR11 endochitinase, Bt serovar kurstaki chitinase (kchi), Bt strain MR21 endochitinase and Bacillus cereus B4264.

In North America, uncultivated, free-living grapevines (Vitus spp.) frequently grow alongside their cultivated counterparts, thus increasing the potential for exchange of microbiota. For this study we used high-throughput sequencing (HTS) of small RNAs (sRNA-Seq) to survey for virus populations in free living grapevines of the Finger Lakes region of New York (NY) State. Of thirty-two grapevines analyzed, twenty-three were free-living vines, while the remaining nine were commercial grown V. vinifera plants from the same region. In total, eighteen (78.3%) of the free-living grapevines tested were positive for grapevine asteroid mosaic-associated virus (GAMaV) infection by HTS, with detection confirmed by semi-nested RT-PCR and sequencing of nine isolates. Phylogenetic analyses of an ungapped alignment of the NY GAMaV sequences (length 2334 nt) with the five known full-length or close to full length global sequences showed the NY isolates were broadly grouped. Of the nine cultivated plants, eight were infected with both hop stunt viroid and grapevine yellow speckle viroid 1, three were singly infected with grapevine leafroll-associated virus 3 and one harbored GAMaV. This limited survey of free-living grapevines, one of the first to use HTS, has highlighted the high incidence of a virus associated with disease in commercial V. vinifera.