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a. eva.ua.ion of ga.lic lectin a. a. a.terna.ive ca.rier doma.n for insecticida. fusion proteins

Insect Science 2008年第6期15卷 483-495页

作者： Ela.ne Fitches Judith Philip Ga.eth Hinchliffe Leisbeth Vercruysse Na.a.a.eb Chougule john a. gatehouse Centra. Science La.ora.ory Sand Hutton York Y041 1LZ UK School of Biologica. a.d Biomedica. Sciences Durham University South Road Durham DH1 3LE UK

The ma.nose-binding lectin GNa.（snowdrop lectin） is used a. a.ca.rier＂ doma.n in insecticida. fusion proteins which cross the insect gut a.ter ora. ingestion. a.simila. lectin from ga.lic bulb, a.a.I, ha. been eva.ua.ed a. a. a.terna.ive ＂ca.rier＂. Recombina.t a.a.I delivered ora.ly to la.va. of ca.ba.e moth （Ma.estra.bra.sica. Lepidoptera. wa. subsequently detected in ha.molymph, demonstra.ing tra.sport. Fusion proteins comprising a. insect neurotoxin, Buta.T （Buthus ta.ulus insecticida. toxin; red scorpion toxin） linked to the C-termina. region of a.a.I or GNa.were produced a. recombina.t proteins （GNa. Buta.T a.d a.a.Buta.T） by expression in Pichia.pa.toris. In both ca.es the C-termina. sequence of the lectin wa. trunca.ed to a.oid post-tra.sla.iona. proteolysis. The GNa. conta.ning fusion protein wa. toxic by injection to ca.ba.e moth la.va. （LD50≈ 250μg/g）, a.d when fed ha. a.nega.ive effect on surviva. a.d growth. It a.so decrea.ed the surviva. of cerea. a.hids （Sitobion a.ena.; Homoptera. from neona.e to a.ult by 〉70% when fed. In contra.t, the a.a.Buta.T fusion protein wa. non-toxic to a.hids, a.d ha. no effect on lepidoptera. la.va., either when injected or when fed. However, inta.t a.a.Buta.T fusion protein wa. present in the ha.molymph of ca.ba.e moth la.va. following ingestion, showing tha. tra.sport of the fusion ha. occurred. The sta.ilities of GNa.Buta.T a.d a.a.Buta.T to proteolysis in vivo a.ter injection or ingestion differed, a.d this ma. be a.fa.tor in determining insecticida. a.tivities.

关键词： a.lium sa.ivum lectin a.a.I gut to ha.molymph tra.sport Pichia.pa.toris expression system sta.ility to proteolysis

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Responses of Whea. (Triticum a.stivum) to Gra.n a.hid (Sitobion a.ena.) Infesta.ion a.d Mecha.ica. Wounding Using a.cDNa.Subtra.titve Libra.y a.proa.h

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a.ricultura. Sciences 2022年第6期13卷 715-740页

作者： Wenzhu Gua. Ma.tin G. Edwa.ds john a. gatehouse a.gha.a. M. R. gatehouse School of Na.ura. a.d Environmenta. Sciences Newcastle University Newcastle UK School of Biologica. a.d Biomedica. Sciences University of Durham Durham UK

a.hids a.e ma.or insect pests of cerea. crops, a.ting a. virus vectors a. well a. ca.sing direct da.a.e. The responses of commercia. whea. (cv. Cla.re) to gra.n a.hid (Sitobion a.ena.) infesta.ion a.d mecha.ica. wounding were investiga.ed in this study, with the a.m to eventua.ly identify a.source of molecula. ma.kers to breed whea. for enha.ced insect resista.ce, a.d in pa.ticula. for enha.ced resista.ce to phloem-feeding insects. Mecha.ica. wounding wa. included in this study a. a.compa.ison with a.hid feeding to distinguish between insect-specific responses in whea. pla.ts to those involved in a.genera. wounding response. Whea. (Triticum spp.) is known to ha.e pa.tia. resista.ce towa.d a.hids [1]. The pla.t response a.d defence a.a.nst insect feeding a.e complica.ed, but a.wa.s follow the sa.e principle: insect detection, signa. tra.smission to initia.e defence, cha.ges in pla.t gene expression a.d subsequent production of defensive compounds, which ma. be ta.geted to the wound site to deter or kill insects. Defensive gene products/proteins rea.h the ta.get a.ea.a.d deter or kill insects. Whether the la.t step is successful or not depends on the resista.ce a.d susceptibility of the pla.t towa.ds tha. pa.ticula. pest. In the light of this principle, it is importa.t to detect cha.ges in gene expression, first a. the tra.scriptiona. level, which is useful for detection of ea.ly-sta.e responses, a.d then once sufficient time is a.lowed for the pla.t to produce defensive gene products, responses a. the proteome level ca. be identified. Work presented in this study focuses on the cha.ges a. the tra.scriptiona. level;differentia. responses a. the proteome level were investiga.ed a.d presented in Ferry et a.. 2011 [2] a.d Gua. et a.. 2015 [3]. Two cDNa.subtra.tive hybridiza.ion libra.ies were constructed, one to identify tra.scripts involved in the responses to a.hid infesta.ion, a.d the second to identify tra.scripts involved in responses to mecha.ica. wounding. Followi

关键词： Whea. Gra.n a.hid Response Resista.ce Tolera.ce cDNa.Subtra.tive Libra.y

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