【作者】 郭广君；

【导师】 杜永臣；

【作者基本信息】 中国农业科学院 ， 蔬菜学， 2013， 博士

【摘要】 番茄（Solanum lycopersicum.Mill）生长过程中易受多种病虫害的影响，属于病虫害多发的蔬菜作物。B型烟粉虱属于世界性入侵害虫，2007年以来烟粉虱危害对我国番茄生产造成了毁灭性的打击，南方露地和北方保护地番茄严重减产甚至绝收。目前无论是国际市场还是国内市场都没有成熟的番茄抗烟粉虱品种。与番茄抗病和抗逆性研究相比，番茄抗虫性机理更为复杂，遗传背景较为薄弱，严重阻碍了抗虫种质资源的利用，抗虫基因的转化和抗虫育种的进展。针对以上问题。本研究以普通番茄(S. lycopersicum)‘9706’、‘LA3556’、‘毛粉802’，潘那利番茄(S. pennellii)‘LA0716’，多毛番茄（S. habrochaites）‘LA2329’、‘LA1777’、‘PI134417’和醋栗番茄(S. pimpinellifolium)‘TO937’、‘PI126933’为试验材料，通过对番茄叶表腺毛类型、密度和次生代谢物质差异分析及与番茄对烟粉虱抗性的相关性分析；明确了不同番茄材料抗虫性状差异，进而确定了番茄抗虫性状与番茄对烟粉虱抗性间的关系；分析接烟粉虱处理后番茄叶表次生代谢物质的变化及相关基因表达量的变化，探讨不同番茄材料对烟粉虱的抗性机制；同时对番茄抗烟粉虱相关QTL进行分析。主要研究结果如下：1.多毛番茄‘LA2329’、‘LA1777’和‘PI134417’和潘那利番茄‘LA0716’对烟粉虱抗性最强；醋栗番茄‘TO937’的抗性具有明显的发育阶段特异性；普通番茄‘9706’、‘LA3556’、‘毛粉802’和醋栗番茄‘PI126933’对烟粉虱无抗性。2.不同番茄材料叶表腺毛类型和密度具有显著的种间和种内差异。抗烟粉虱的多毛番茄LA1777、LA2329、PI134417，潘那利番茄LA0716和醋栗番茄TO937叶表密布有腺体的Ⅳ型腺毛，其密度＞5个·mm-2；不抗烟粉虱的普通番茄9706、LA3556、毛粉802及醋栗番茄‘PI126933’叶表主要为无腺体的Ⅲ型腺毛，密度为3.4~10.6个·mm-2。此外，3份多毛番茄叶表Ⅵ型腺毛密度显著高于其他番茄材料。叶表Ⅳ型腺毛密度和萜类物质含量与叶表烟粉虱产卵量显著负相关；Ⅵ型腺毛密度与叶表萜类物质含量显著正相关，与烟粉虱产卵量无相关性；叶表无腺体的Ⅲ型腺毛密度与烟粉虱产卵量显著正相关。3.本研究中，抗烟粉虱的3份多毛番茄叶表萜类物质的种类和含量高于其他番茄材料，同时具有显著的种内差异。‘LA1777’叶表萜类物质共有24种，包括3种单萜类物质和21种倍半萜类物质，其中CⅠ类和CⅡ类倍半萜类物质含量差异不大。‘LA2329’的萜类物质共有24种，包括5种单萜和19种倍半萜类物质，主要为CⅡ类倍半萜和与之结构相同的倍半萜类物质。‘PI134417’中萜类物质为14种，包含1种单萜和13种倍半萜类物质，主要为CⅠ类倍半萜和与之结构相同的倍半萜类物质。抗烟粉虱的潘那利番茄‘LA0716’和醋栗番茄‘TO937’与不抗烟粉虱的普通番茄‘9706’、‘LA3556’、‘毛粉802’和醋栗番茄‘PI126933’叶表共检测到10种萜类物质，包括6种单萜和4种CⅠ类倍半萜。不同发育时期，番茄叶表萜类物质种类变化较小，物质含量差异显著。4.接烟粉虱处理后，多毛番茄‘LA1777’共有21种倍半萜类物质具有显著差异。接虫8h、48h和21d处理后，分别有3种、8种和10种萜类物质其含量显著增加。‘LA2329’共有12种倍半萜类物质具有显著差异。接虫8h和48h处理后，含量较高的7种物质，含量显著降低，4种微量的物质，含量显著增加；接虫21d处理后，11种物质含量均显著增加。‘PI134417’只有2种倍半萜类物质具有显著差异。接虫48h后，物质含量显著降低，接虫21d后，含量显著增加。5.萜类合成途径的基因表达分析显示，3份多毛番茄材料萜类物质合成途径中基因表达量显著高于其他番茄材料，说明番茄可在转录水平调控倍半萜类物质的合成。接烟粉虱处理后，基因表达差异最大的为前体物质FPP合酶基因FPS和萜类合酶基因TPS9、TPS12和SSTLH3，其基因表达量与倍半萜类物质含量基本一致，说明这几个基因为调控萜类物质含量积累的主要基因。6.利用多毛番茄‘LA2329’和普通番茄‘9706’构建的BC1群体中，共定位到33个与番茄抗烟粉虱相关的QTL。叶片烟粉虱成虫附着量和产卵量定位的QTL位点相同，共定位到8个QTL，其中2个主效QTL定位在2号染色体，分别介于标记InDel_FT45~SSR57之间和SSR57~InDel_FT49之间，解释的表型变异率分别为33.2%和39.8%。更多还原

【Abstract】 A number of arthropod pests attack tomato throughout its production cycle. Bemisia tabaci(Gennadius) biotype B is a worldwide pest and since2007has become a severe pest in China causingextreme damage to field tomato production in southern China and to greenhouse tomato production innorthern China. There are no tomato varieties resistant to whitefly in the international and domesticmarkets. Compared with disease and abiotic stress resistance mechanisms, tomato insect resistancemechanisms are more complex and in only a few instances have the genetic basis for resistance beeninvestigated. These problems prevent use of insect-resistant germplasm resources, transformation ofinsect resistance genes and development of breeding for insect resistance.S.lycopersicum ‘9706’,‘LA3556’,‘maofen802’, S.habrochaites ‘LA2329’,‘LA1777’,‘PI134417’,S. pennellii ‘LA0716’and S. Pimpinellifolium ‘TO937’,‘PI126933’ were used in this study. In order toillustrate the mechanism of different tomato accessions resistance to Bemisia tabaci. This reasearchinvolved correlational analysis for trichomes, secondary metabolites on leaf surface and resistance towhitefly among the tomato accessions; secondary metabolite profiles on the leaf surfaces, geneexpression resulting from several whitefly treatments and identification of QTL‘s associated withresistance to Bemisia tabaci in S. habrochaites LA2329. Main results for this study are as follows:1. The resistance of S.habrochaites ‘LA2329’，‘LA1777’，‘PI134417’and S. pennellii ‘LA0176’towhitefly was the strongest. The resistance of S. pimpinellifolium ‘TO937’ to whitefly was significantdifference in different developmental stages. S.lycopersicum ‘9706’,‘LA3556’,‘maofen802’ and S.Pimpinellifoliu ‘PI126933’ were sensitive to whitefly.2. Trichome type and density on abaxial leaf surfaces of the different tomato accessions showedsignificant inter-and intra-specific differences. Type Ⅳ and Ⅵ trichome densities were superior onabaxial leaf surfaces of S. habrochaites, the densities of them were more than5·mm-2. Type Ⅳ trichomedensities were superior on abaxial leaf surfaces of S. pennellii and S. pimpinellifolium ‘TO937’, thedensities were6.7and15.4·mm-2, respectively. Other tomato accessions trichomes on abaxial leafsurfaces were mainly non-glandular trichomes, densities were3.4~10.6·mm-2. Type Ⅳ and Ⅵ trichomedensities were less than4·mm-2on abaxial leaf surface of these tomato accessions. Density of type Ⅳtrichomes, wax content on leaf surfaces, and terpenoid content were significantly negative correlatedwith number of whitefly eggs laid on abaxial leaf surfaces. Conversely, density of type Ⅲ trichomeswas positive correlated with the number of whitefly eggs laid on abaxial leaf surfaces. Density of typeⅥ trichome was positive correlated with terpenoid content, but not correlated with number of whiteflyeggs laid on abaxial leaf surfaces.3. Terpenoids of S. habrochaites ‘LA2329’,‘LA1777’ and ‘PI134417’ were mainly sesquiterpenes,which showed significant inter-and intra-specific differences. Terpenoids of ‘LA1777’ were25compositions including3monoterpenes and22sesquiterpenes that comprised of type CⅠand CⅡsesquiterpenes. Terpenoids of ‘LA2329’ were24compositions including5monoterpenes and16 sesquiterpenes that comprised of type CⅡ sesquiterpenes and sesquiterpenes with the same chemicalstructure of type CⅡ sesquiterpenes. Terpenoids of ‘PI134417’ were14compositions including1monoterpenes and13sesquiterpenes that comprised of type CⅠ sesquiterpenes and sesquiterpenes withthe same chemical structure as type CⅠ sesquiterpenes. Conversely，terpenoids of S. lycopersicum, S.pennellii and S. pimpinellifolium were mainly monoterpenes, which were10compositions andcomprised of6monoterpenes and4type CⅠ sesquiterpenes, which showed little diversity among thethree tomato species. Terpenoid composition was relatively small different among developmental stages,but terpenoid concentration differed significantly among developmental stages.4. Secondary metabolites were significantly different among3tomato accessions of S.habrochaitesafter whitefly treatment,compared to uninfested controls. A total of21kinds of sesquiterpenes of‘LA1777’ were significant difference after whitefly treatment. After8h by whitefly adults infestion,48h exposure to whitely eggs and21d by whitefly nymphs infestion,3sesquiterpenes,8sesquiterpenesand10sesquiterpenes significantly increased. A total of12kinds of sesquiterpenes of ‘LA2329’ weresignificant difference after whitefly treatment. After8h by whitefly adults infestion and48h exposureto whitely eggs7sesquiterpenes significantly decreased and4sesquiterpenes significantly increased.After21d by whitefly nymphs infestion,11sesquiterpenes significantly increased. A total of2kinds ofsesquiterpenes of ‘PI134417’ were significant difference after whitefly treatment. After8h by whiteflyadults infestion and48h exposure to whitely eggs these sesquiterpenes significantly decreased, andafter21d by whitefly nymphs infestion, significantly increased.5. Expression of genes associated with tomato terpenoid biosynthesis of S. habrochaites‘LA2329’,‘LA1777’and ‘PI134417’were highter than others tomato accessions. The result illustrategenes control sesquiterpenes biosynthesis in the transcriptional level. The terpenoid gene expression oftomato accessions was not significant affected by treatment of adult whiteflies. Expressions of FPS,TPS9, TPS12and SSTLH3of tomato accessions were significantly difference after whitefly treatment,compared to uninfested controls. This suggest FPS，TPS9，TPS12and SSTLH3were the key geneswhich regulate sesquiterpenes biosynthesis.6.33QTL associated with insect-resistance were mapped in the BC1population generated fromtomato cultivar‘9706’crossed with S. habrochaites‘LA2329’.8QTL associated with adult whiteflynumbers and oviposition numbers were located in the same locus. Of the8QTL,2major QTL werelocated in the interval defined by the markers InDel_FT45~SSR57and SSR57~InDel_FT49onchromosome2and explained33.2%and39.8%of phenotypic variation，respectively.更多还原