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中国小桐子不同地理种源差异比较与种源亲缘关系研究

Studies on Genetic Relationship of Provenance and Comparative Analysis of Provenance Difference of Jatropha Curcas Linn

【作者】 栗宏林

【导师】 张志翔; 林善枝;

【作者基本信息】 北京林业大学 , 野生动植物保护与利用, 2008, 博士

【摘要】 随着能源危机的日益迫近,环境友好、高效可再生的生物质能源的研究开发正方兴未艾。小桐子以高含油率、优质的特点成为最有发展潜力的生物能源树种之一。本研究对我国小桐子主要分布区内22个种源从形态学、解剖学、光合生理三个层次对不同地理种源间差异进行比较研究,并采用ITS测序对不同种源的小桐子亲缘关系与起源进行探讨。研究结果丰富了小桐子系统进化学研究的内容,为进一步深入探讨小桐子亲缘关系提供依据;同时对实际生产中种子的调拨、优良种源的选择育苗及造林生产、扩大种植区等工作提供理论依据,增加资源产量,解决生物质能源开发原料不足带来的“瓶颈”问题,具有重要实际意义。主要结论如下:1)形态学研究表明不同种源地种子形态性状差异明显,表明小桐子不同种源遗传多样性丰富,种源选择具有很大潜力。不同种源出苗率及株高存在显著差异,种源间地径无显著差异。综合种子性状及苗期生长,初步提出生长表现较好的种源,可作为进一步开展地理种源造林试验和优良种源推广的材料。2)叶片解剖观察表明,小桐子幼苗叶片具有双子叶植物典型的异面叶解剖结构。其叶片横切面由上表皮、叶肉组织、叶脉及下表皮四个部分组成。小桐子不同种源地幼苗叶片的各部组成结构相同,但各结构部分的紧密度及厚度具有明显不同。小桐子22个种源间叶片厚度、上表皮厚度、栅栏组织厚度、海绵组织厚度、栅海比、叶片结构紧密度及叶片结构疏松度等解剖性状存在显著差异。解剖结构性状彼此间存在不同程度的相关性。株高与叶片上表皮厚度极显著正相关,而与叶片结构紧密度呈显著负相关;小桐子幼苗生长势仅表现为株高与叶片结构有一定相关性。3)小桐子一年生苗木光合速率日变化呈双峰型,有“光合午休”的现象。光合速率年周期变化分为“单峰型”和“双峰型”两种,单峰型种源多分布于纬度较低的区域,双峰型种源多分布于纬度略高的山地地区。不同种源小桐子光合速率在一年的生长季中各不相同,但光合速率有随纬度升高呈现增加的趋势。不同种源叶绿素含量、气孔导度(Cond)、胞间CO2浓度(Ci)、蒸腾速率(Trmmol)及光饱和点、光补偿点等光合特征各不相同,存在种源选择的可能。4)经ITS序列分析,小桐子各种源间存在变异,遗传多样性较丰富。小桐子不同种源的ITS序列全长为479~649bp,ITS1的长度范围为163~278bp,ITS2的长度范围为116~207bp,5.8S FDNA为155bp~227bp。G+C的含量在ITS1区的变化范围为34.7%~66.2%,在ITS2区的变化范围为34.1%~69.5%。所有样品中有12个样品的ITS全序列的长度高度一致,均为648bp,其中有142个变异位点占ITS的21.91%。ITS1共有278bp,其中有70个变异位点,占ITS1的25.18%;ITS2共有207 bp,其中有46个变异位点,占ITS2的22.22%。5)小桐子18个种源由系统发生树分为4个组,亲缘关系远近不同,种源来源不同,表明我国的小桐子是经由多条路径引种而来而非同一来源。亲缘关系的系统分组与形态学分组不完全符合。分布于我国云南省的8个地区的种源具有同一起源;广西省8个地区的种源具有不同的来源。

【Abstract】 With the energy crisis increasingly urgent, the research and development of environment-friendly, high efficiency and renewable bio-energy is on the up. Due to high oil content and high quality, Jatropha curcas L.became one of the most potential bio-energy plant. In this paper about twenty-two Jatropha curcas provenances in its primarily distributes area were comparatively study on morphology, anatomy and photosynthesis. Also adapt ITS mensurated order to discuss genetic relationship and origin of different Jatropha curcas L.provenances. The results enriched the content of phylogenetics and offered gist for discuss further the genetic relationship between different provenances; At the same time provided academic bases for seed transfer, eminent provenance’s breeding, enlarge plant region and so on . Further more there were important practical meanings in increasing output, solving the bottle-neck problem because of scarcity material. Main conclusions were as follows:1) The results showed that morphologic characteristics of seeds for different provenances had significant variation. This indicated Jatropha curcas L.of different provenances had abundant genetic diversity and the selection of provenances had big potential. Different provenances had significant variation in seedling emergence percentage and plant height, ground diameter were not Synthesizing seed character and seedling periods grows, preliminary put forward the provenances of good growth performance , this can be used as the material to futher open forestation experiment of geography provenances and the extend of eximious provenances.2) The observed results of leaf anatomical structure indicated that the leaf of Jatropha curcas L. belongs to typical bifacial leaf of dicotyledon. Leaf anatomical structure was composed of four parts: upper epidermis, mesophyll tissue, vein and lower epidermis . Each part of seedling leaves anatomical structure of Jatropha curcas L. from different provenances was same to others, but there were significant differences in tightness and thickness of each part. The results showed that there were significant differences in leaf thickness, thickness ratio between palisade tissue and spongy tissue, the thickness of upper epidermis, cell tense ratio(CTR) and spongy ratio(SR) of different provenances. Characters were related to each other at different degree. There were positive correlation between seedling height and upper epidermis, while there was negative correlation between seedling height and cell tense ratio (CTR). Correlation between growth vigor of seedling and leaf anatomical structure mainly shows in seedling height was related to leaf structure.3) The results indicated that diuilial variation of Pn was double peak curve in seedling of Jatropha curcas L., and the annual change pattern of Pn of Jatropha curcas L. are divided into two types, double peak curve type and single peak type. Provenances of single peak type mainly distributed in low-latitude region, while provenances of double peak type mainly distributed in high-latitude mountain area. The Pn of Jatropha curcas L. of different provenances was obciously different, it increased with the increasing of latitude. The results showed that the different photosynthetic parameters of different provenances were distinct, which indicated high provenances selection potential in the different provenances.4) The sequence of the ITS (including ITS1, 5. 8SrDNA and ITS2) of nuclear ribosomal DNA from Jatropha curcas L. from 22 provenances were determined. The results showed that the size of the ITS of Jatropha curcas L. was from 479 to 649 bp (ITS1 is from 163 to 278 bp, ITS2 is from 116 to 207bp). The length of 5.8S rDNA was from 155bp to 227 bp. The range of G+C contene of ITS1 was variated from 34.7% to 66.2%, while the contene of G+C in ITS2 was from 34.1%~69.5%. The size of the ITS of 12 sample was very conservative, its length was 648bp.It was found that there were 142 varied characters, the percentage of varied characters was 21.91%. In addition, there were 70 varied characters in total 278 characters of ITS1, its percentage was 25.18%. There were 46 varied characters in total 207 charactersof ITS2, its percentage was 22.22%.5) The results showed that Jatropha curcas L. from 18 different provenances were divided four groups by Means of phylogenetic tree analysis. It indicateed that phylogenetic relationship of Jatropha curcas L. from 18 different provenances was different. It proved that Jatropha curcas L. of China was introducted through several routes. Phylogram tree based on ITS sequence data indicated that the samples from same region was not always clustered, and the samples from different regions may be in same section. This was not corresponding to the result that determined by morphology. It was found that Jatropha curcas L. of Yunnan Province from same source, but which from Guangxi Province was origin multiple.

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