The tea mosquito bug (TMB), Helopeltis spp. (Hemiptera: Miridae) is an insidious pest that poses a significant economical threat to tea plantations. Pseudomonas cultures are being used extensively for pest management which, however, resulting in a low mortality rate of insects and which has prompted us to search for a new microbial metabolite for TMB control. A chitinase purified from P. fluorescens and partially characterized by our group showed insecticidal activity against TMB. The mode of action behind chitinase toxicity is the enzymatic hydrolysis of chitin, which is a common constituent of the insect exoskeleton and gut lining of the peritrophic membrane. A chitinase-secreting strain MP-13 was characterized based on 16S rRNA sequencing and validated as Pseudomonas fluorescens. In the present study, purified chitinase (0.048 units/ml) enzyme from P. fluorescens MP-13 revealed 100% TMB mortality under in-vitro conditions. The results of this study can be utilized for future crop improvement programs and integrated pest management strategies.

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Background Cotton is one of the most important commercial crops as the source of natural fiber, oil and fodder. To protect it from harmful pest populations number of newer transgenic lines have been developed. For quick expression checks in successful agriculture qPCR (quantitative polymerase chain reaction) have become extremely popular. The selection of appropriate reference genes plays a critical role in the outcome of such experiments as the method quantifies expression of the target gene in comparison with the reference. Traditionally most commonly used reference genes are the “house-keeping genes”, involved in basic cellular processes. However, expression levels of such genes often vary in response to experimental conditions, forcing the researchers to validate the reference genes for every experimental platform. This study presents a data science driven unbiased genome-wide search for the selection of reference genes by assessing variation of > 50,000 genes in a publicly available RNA-seq dataset of cotton species Gossypium hirsutum. Result Five genes (TMN5, TBL6, UTR5B, AT1g65240 and CYP76B6) identified by data-science driven analysis, along with two commonly used reference genes found in literature (PP2A1 and UBQ14) were taken through qPCR in a set of 33 experimental samples consisting of different tissues (leaves, square, stem and root), different stages of leaf (young and mature) and square development (small, medium and large) in both transgenic and non-transgenic plants. Expression stability of the genes was evaluated using four algorithms - geNorm, BestKeeper, NormFinder and RefFinder. Conclusion Based on the results we recommend the usage of TMN5 and TBL6 as the optimal candidate reference genes in qPCR experiments with normal and transgenic cotton plant tissues. AT1g65240 and PP2A1 can also be used if expression study includes squares. This study, for the first time successfully displays a data science driven genome-wide search method followed by experimental validation as a method of choice for selection of stable reference genes over the selection based on function alone.

Prevalence of Races and Biotypes ofRalstonia Solanacearumin IndiaBacterial wilt caused byRalstonia solanacearumis the most destructive disease of plants. Fifty-seven isolates ofR. solanacearumcausing wilt on different host plantsviz., tomato (Solanum lycopersicum), brinjal (S. melongena), potato (S. tuberosum), bird of paradise (Strelitzia reginae), ginger (Zingiber officinale), chili (Capsicum annuum), capsicum (Capsicum annuum), davana (Artemisia pallens) and coleus (Coleus forskohlii) were collected from the different agro climatic zones of Karnataka and other parts of India. In this study, 57 isolates were differentiated into race on the basis of their pathogenicity and their ability to infect different host. The isolates were established as race-1. None of the isolates infected mulberry and banana. Fifty-four isolates oxidized and utilized both the disaccharides and sugar alcohols. These isolates were positioned as biovars-3 according to Haywards classification system. Three isolates from Kerala, two ginger, and one tomato strain were not able to utilize dulcitol and lactose. Hence, they were categorized into a new taxo group within the system and designated as biovar-3B for the first time in India. There were 54 isolates which were confirmed as race-1, biovar-3, and 3 isolates were confirmed as race-1, biovar-3B by morphological, physiological, biochemical and pathogenicity studies. Two sets of primers (OLI1 & Y2 and Y1 & Y2) were used in this study to authenticate the organism. Furthermore, the identity of the isolates was confirmed by a serological diagnostic kit obtained from the International Potato Research Center, Lima, Peru, and single chain variable fragment antibody specific toR. solanacearum.