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In vitro and in vivo application of RNA interference for targeting genes involved in peritrophic matrix synthesis in a lepidopteran system

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Insect Science 2013年第1期20卷 92-100页

作者： Umut Toprak Doug Baldwin Martin Erlandson Cedric Gillott stephanie harris Dwayne D. Hegedus Agriculture and Agri-Food Canada Ottawa Ontario Canada Department of Biology University of Saskatchewan Canada Department of Plant Protection Faculty of Agriculture University of Ankara Turkey Department of Food and Bioproduct Sciences University of Saskatchewan Saskatoon SK Canada

The midgut of most insects is lined with a semipermeable acellular tube, the peritrophic matrix （PM）, composed of chitin and proteins. Although various genes encoding PM proteins have been characterized, our understanding of their roles in PM structure and function is very limited. One promising approach for obtaining functional information is RNA interference, which has been used to reduce the levels of specific mRNAs using double-stranded RNAs administered to larvae by either injection or feeding. Although this method is well documented in dipterans and coleopterans, reports of its success in lepidopterans are varied. In the current study, the silencing midgut genes encoding PM proteins （insect intestinal mucin 1, insect intestinal mucin 4, PM protein 1） and the chitin biosynthetic or modifying enzymes （chitin synthase-B and chitin deacetylase 1） in a noctuid lepidopteran, Mamestra configurata, was examined in vitro and in vivo. In vitro studies in primary midgut epithelial cell preparations revealed an acute and rapid silencing （by 24 h） for the gene encoding chitin deacetylase 1 and a slower rate of silencing （by 72 h） for the gene encoding PM protein 1. Genes encoding insect intestinal mucins were slightly silenced by 72 h, whereas no silencing was detected for the gene encoding chitin synthase-B. In vivo experiments focused on chitin deacetylase 1, as the gene was silenced to the greatest extent in vitro. Continuous feeding of neonates and fourth instar larvae with double-stranded RNA resulted in silencing of chitin deacetylase 1 by 24 and 36 h, respectively. Feeding a single dose to neonates also resulted in silencing by 24 h. The current study demonstrates that genes encoding PM proteins can be silenced and outlines conditions for RNA interference by per os feeding in lepidopterans.

关键词： chitin deacetylase chitin synthase-B lepidoptera peritrophic matrix,peritrophin RNA interference

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X-ray imaging of 30 year old wine grape wood reveals cumulative impacts of rootstocks on scion secondary growth and Ravaz index

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Horticulture Research 2023年第1期10卷 31-44页

作者： ZoëMigicovsky Michelle Y.Quigley Joey Mullins Tahira Ali Joel F.Swift Anita Rose Agasaveeran Joseph D.Dougherty Brendan Michael Grant Ilayda Korkmaz Maneesh Reddy Malpeddi Emily L.McNichol Andrew W.Sharp Jackie L.harris Danielle R.Hopkins Lindsay M.Jordan Misha T.Kwasniewski R.Keith Striegler Asia L.Dowtin stephanie Stotts Peter Cousins Daniel H.Chitwood Plant Foodand Environmental SciencesFaculty of AgricultureDalhousie UniversityTruroNova ScotiaB2N 5E3Canada Department of Horticulture Michigan State UniversityEast LansingMI48823USA College of Natural Science Michigan State UniversityEast LansingMI48823USA Department of Neuroscience Michigan State UniversityEast LansingMI48823USA Department of Biology Saint Louis UniversitySt.LouisMO63103USA Department of Physics and Astronomy Michigan State UniversityEast LansingMI48823USA Department of Computational Mathematics Science&EngineeringMichigan State UniversityEast LansingMI48823USA College of Engineering Michigan State UniversityEast LansingMI48823USA College of Social Science Michigan State UniversityEast LansingMI48823USA Department of Economics Michigan State UniversityEast LansingMI48823USA Department of Biochemistry and Molecular Biology Michigan State UniversityEast LansingMI48823USA College of Arts and Letters Michigan State UniversityEast LansingMI48823USA E.&J.Gallo Winery AcampoCA95220USA Current affiliation:Constellation Brands SoledadCA93960USA Department of Food Science The Pennsylvania State UniversityState CollegePA16803USA Department of Forestry Michigan State UniversityEast LansingMI48823USA Department of Agriculture Foodand Resource SciencesUniversity of Maryland Eastern ShorePrincess AnneMD21853USA Department of Natural Sciences University of Maryland Eastern ShorePrincess AnneMD21853USA E.&J.Gallo Winery ModestoCA95354USA Department of Biology Acadia UniversityWolfvilleNova ScotiaB4P 2R6Canada

Annual rings from 30 year old vines in a California rootstock trial were measured to determine the effects of 15 different rootstocks on Chardonnay and Cabernet Sauvignon *** traits measuring vegetative growth,yield,berry quality,and nutrient uptake were collected at the beginning(1995 to 1999)and end(2017 to 2020)of the lifetime of a vineyard initially planted in 1991 and removed in 2021.X-ray Computed Tomography(CT)was used to measure ring widths in 103 *** width was modeled as a function of ring number using a negative exponential *** and late wood ring widths,cambium width,and scion trunk radius were correlated with 27 *** of annual ring width shows that scions alter the width of the first rings but that rootstocks alter the decay of later rings,consistently shortening ring width throughout the lifetime of the *** index,juice pH,photosynthetic assimilation and transpiration rates,and instantaneous water use efficiency are correlated with scion trunk ***,our research indicates that rootstocks modulate secondary growth over years,altering physiology and agronomic *** act in similar but distinct ways from climate to modulate ring width,which borrowing techniques from dendrochronology,can be used to monitor both genetic and environmental effects in woody perennial crop species.

关键词： rings lifetime removed

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