巨桉重要性状GWAS分析及抗桉树枝瘿姬小蜂整合研究

发布时间：2018-05-30 18:16

本文选题：巨桉 + 重要性状　；参考：《中国林业科学研究院》2017年博士论文

【摘要】：巨桉天然分布新南威尔士纽卡斯尔、昆士兰中部班德堡以及昆士兰北部麦凯、汤斯维尔和丹特里等沿海或近海岸地区,因其广泛的适应性和优良加工利用特性,成为桉树育种者首选育种材料。论文以2010年栽培的巨桉8个种源实验林为研究材料,根据核心种质库构建理论,筛选主要性状可以稳定变异的100株为最终研究对象。系统地开展生长量、干形评价、叶片特征、木材材性、桉树枝瘿姬小蜂危害调查及评价、桉树枝瘿姬小蜂危害后相关酶活性测定、叶片含糖量、叶片挥发性化合物含量测定等研究;结合巨桉基因组高通量测序数据,进行性状的全基因组关联分析;利用桉树枝瘿姬小蜂危害调查评价材料,筛选3株高感、3株高抗无性繁殖材料,进行桉树枝瘿姬小蜂抗性转录组研究;整合巨桉生理生化代谢产物和转录组数据,全面探索桉树枝瘿姬小蜂抗性机理。主要研究结论如下:1.巨桉生长量种源间差异不显著,6年生树高15.00~17.02m、枝下高6.83~8.70m、胸径11.28~14.21cm、单株材积0.08~0.15m3、冠幅1.56~1.86m、通直度3.30~3.86、圆满度2.20~3.21、分枝1.60~2.86、分枝角度47.69~62.50°;单株之间差异显著,单株选择有效,主成分分析筛选最优良单株74号,得分1.6。2.巨桉种源基本密度在0.38~0.44g.cm-3范围,平均0.42g.cm-3;导管比量在10.06-12.18%范围,平均11.00%;薄壁组织比量在2.46%~2.55%范围,平均2.51%;木射线比量在12.89~14.73%范围,平均13.84%;木材纤维宽度在7.39~7.90μm范围,平均7.62μm;木材纤维长度在891.81~976.36μm范围,平均932.95μm;材纤维长宽比在107.02~181.40范围,平均124.77;木材微纤丝角在16.22~18.48°范围,平均17.42°。木材基本密度、薄壁组织比量、木纤维、纤维宽度、纤维长度、纤维长宽比均表现由髓心向树皮处逐渐增大;薄壁组织比量、木射线比量和微纤丝角由髓心向树皮处逐渐减小。3.桉树枝瘿姬小蜂危害调查,高感种源16892、17907、20555、20674各有2株,种源18146、20261和South Africa各1株,种源18705有4株;高抗单株41株。4.防御性酶在不同种源含量有差别,但不显著;单株间PPO、CAT、POD酶活力差异显著,SOD酶活力差异不显著。叶绿素b含量在种源间和单株间差异均不显著,叶绿素a和类胡萝卜素含量在种源间差异不显著;单株间差异显著。5.桉树叶挥发性成分主要成分单萜类化合物,化合物相对含量均在80%以上;单萜醇酯相对含量变化显著,在6.27~11.79%范围;种源18146、18705、20261和南非的挥发性成分—单萜醇酯相对含量在10%以上。17号单萜类化合物相对含量最大,达96.87%,主成分是α-蒎烯(57.94%)、γ-松油烯(22.80%)和对伞花烃(10.46%);96号单萜醇酯相对含量最多,达44.39%,主要成分是桉叶素(42.20%)、α-蒎烯(31.68%)和对伞花烃(16.52%)。6.巨桉主要性状GWAS分析,与桉树枝瘿姬小蜂危害关联的位点有15个SNP、51个基因定位在8条染色体框上,候选基因注释与植物抗性、适应胁迫环境、病害防御、呼吸及光合作用等有关;与枝下高关联到位点3个;比叶面积关联到1个位点;冠幅关联到6个位点;木材纤维比量关联位点有4个。7.桉树枝瘿姬小蜂抗性转录组研究,发现579个差异表达基因,高感品系与高抗品系相比,360个基因发生上调,219个基因表达下调。GO富集得到244个差异表达基因,富集基因最多的是防御反应相关基因,包含有32个差异表达基因。GO富集层次分析中分子功能富集最显著的8个节点分别为蛋白结合、倍半萜烯合酶活性、泛素蛋白连接酶结合、鸟苷酸环化酶活性、酶抑制剂的活性、组蛋白甲基转移酶活性、月桂烯合酶活性和罗勒烯合酶活性。8.桉树枝瘿姬小蜂整合研究,发现一种脂质转移酶(VAS)在高抗品系内显著表达下调,与黄瓜、辣椒研究结果一致。桉树TPS基因家族整理共包含92个成员,与拟南芥相比发生了显著扩张。高抗品系与高感品系相比,有13个EgrTPS基因的表达显著上调,14个基因的表达显著下调。这些基因可能与桉树抗虫性的差异密切相关,但其功能还有待进一步的研究。
[Abstract]:The natural distribution of Eucalyptus grandis in New South Wales Newcastle, Bundaberg in central Queensland, and the coastal or near coastal areas of Mackay, Townsville and Danli, north of Queensland, has become the first choice breeding material for Eucalyptus breeders because of its extensive adaptability and excellent processing and utilization characteristics. In this paper, the experiment of Eucalyptus grandis in 2010 was the 8 provenance experiment of Eucalyptus Lin Weiyan. According to the construction theory of core germ plasm bank, 100 strains with stable variation can be selected as the final research object. Growth, dry shape evaluation, leaf characteristics, wood properties, damage investigation and evaluation of Eucalyptus gall wasp, determination of related enzyme activity, leaf sugar content and leaf volatilization after the harm of eucalyptus tree gall wasp were systematically carried out. Combined with high throughput sequencing data of Eucalyptus grandis genome, the whole genome association analysis was carried out with high throughput sequencing data of Eucalyptus grandis genome, and 3 high sensitivity and 3 highly resistant reproductive materials were screened by using Eucalyptus clad gall wasp hazard investigation and evaluation materials, and the resistance transcriptional group of eucalyptus tree gall wasp was studied, and the physiological and biochemical metabolism of Eucalyptus grandis was integrated. The main research conclusions are as follows: 1. the growth of Eucalyptus urophylla is not significant, the height of the growth of Eucalyptus grandis is not significant, the height of the tree is 15.00~17.02m, the height of the branch is 6.83~8.70m, the diameter of the breast is 11.28~14.21cm, the individual volume 0.08~0.15m3, the crown 1.56~1.86m, the straightness 3.30~3.86, the satisfactorily 2.20~3.21, branch 1.60~2.86, and the difference. The branch angle 47.69~62.50 degree was significant. The single plant selection was effective. The principal component analysis was used to select the best single plant No. 74. The basic density of the source of Eucalyptus grandis 1.6.2. was in the 0.38~0.44g.cm-3 range, the average 0.42g.cm-3; the catheter ratio was in the 10.06-12.18% range, with an average of 11%; the weight of the thin wall tissue was in the 2.46%~2.55% range, the average 2.51%; the wood ray ratio. The amount is in the range of 12.89~14.73%, with an average of 13.84%; the width of wood fiber is in the range of 7.39~7.90 mu m, with an average of 7.62 m; the length of wood fiber is in the range of 891.81~976.36 mu m, with an average of 932.95 mu m; the ratio of fiber length to width is 124.77, and the Wood Microfibril Angle is in the range of 16.22~18.48 degree, averaging 17.42 degrees, the basic density of wood and the specific parenchyma of the wood. Wood fiber, fiber width, fiber length, fiber length and width ratio increased gradually from pith to bark, and the ratio of parenchyma tissue, wood ray ratio and microfibril angle decreased from the pith to bark of.3. Eucalyptus. There were 2 strains of high sensitivity 16892179072055520674, 1 strains of seed 1814620261 and South Africa, There were 4 plants in 18705 species, and 41.4. resistant enzymes of high resistance single plant were different, but not significant. The activity of PPO, CAT, and POD was not significant, and the difference of SOD enzyme activity was not significant. The content of chlorophyll b was not significant between the sources and among the single plant. The difference of chlorophyll a and carotenoid content was not significant in the species source. The volatile components of.5. eucalyptus leaves were the main components of monoterpenoids, the relative content of the compounds were above 80%, and the relative content of monoterpene esters changed significantly in the 6.27~11.79% range; the relative content of the volatile components of the species 181461870520261 and South Africa, monoterpene ester, was the largest, and the relative content of the.17 monoterpenes was the largest, reaching 96. .87%, the main components are alpha pinene (57.94%), gamma pine oleene (22.80%) and parachute hydrocarbon (10.46%), and 96 mono terpterol ester (96), the main components are Eucalyptus (42.20%), alpha pinene (31.68%) and parachute hydrocarbon (16.52%) of Eucalyptus urophylla, which are related to the loci of the Eucalyptus gall wasp, 15 SNP and 51 genes. On the 8 chromosome frame, the candidate gene annotation and plant resistance, adaptation to stress environment, disease defense, respiration and photosynthesis, are related to the height of the branch height of 3; the specific leaf area is associated with 1 loci; the crown is associated with 6 loci; the wood fiber ratio site has 4.7. Eucalyptus crown gall wasp resistance transcriptional groups. 579 differentially expressed genes were found. Compared with high resistant strains, 360 genes were up-regulated and 244 genes were enriched by 219 down-regulation.GO. The most enriched genes were the defense response related genes, including the 8 nodes of 32 differentially expressed genes in the.GO enrichment level analysis. The integration of protein binding, half terpenoid synthase activity, ubiquitin protein ligase binding, guanosine cyclase activity, enzyme inhibitor activity, histone methyltransferase activity, laurenene synthase activity and.8. Eucalyptus clad gall wasp were integrated, and a kind of lipid transferase (VAS) was found to be expressed in high resistance strains. Down regulation was consistent with the results of cucumber and pepper studies. The TPS gene family of Eucalyptus contains 92 members. Compared with Arabidopsis, there were significant expansion. Compared with high susceptible strains, the expression of 13 EgrTPS genes was significantly up-regulated and the expression of 14 genes decreased significantly. These genes may be closely related to the difference of Eucalyptus resistance. But its function remains to be further studied.
【学位授予单位】：中国林业科学研究院
【学位级别】：博士
【学位授予年份】：2017
【分类号】：S792.39;S763.7

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