【作者】 代莉；

【导师】 刘震；

【作者基本信息】 河南农业大学 ， 森林培育， 2014， 博士

【摘要】 本文通过对江西赣州、贵州贵阳、贵州遵义、江西南昌、湖南张家界、江西九江、浙江临安、湖北宜昌、四川成都、四川广元、重庆、河南洛阳共12个分布区的山桐子种实品质差异性、种子休眠差异性、遗传多样性及生态地理相关性分析，从而得出山桐子种实的地理变异，主要结果如下：（1）12个分布区山桐子果肉含油率达28.38%～48.35%，平均37.62%，种子含油率12.6%～28.17%，平均21.17%，果实（种子+果肉）平均含油率达31.36%；山桐子种子碘值为112.6～139.7mg·g-1，皂化值为191.2～194.6mg·g-1，十六烷值为43.2～49.3mg·g-1，果肉的碘值为108.2～137.1mg·g-1，皂化值为196.2～205.6mg·g-1，十六烷值为43.3～55.0mg·g-1，根据生物柴油产品评价标准，山桐子果实是一种优质的生物柴油原料。其中贵阳分布区山桐子果实最适宜作为生物质柴油原料，可以作为山桐子生物质能源林优先发展区域。洛阳和赣州分布区次之。山桐子果实品质的差异与各分布区的气候条件有一定的相关关系，主要影响因素为有效降雨量、有效日照时数。（2）山桐子果实油脂脂肪酸高达12种（种子10种，果肉12种），本实验测出的9-羟基壬酸、十二酸、十七酸为前人未发现，而亚麻酸、十六碳烯酸在本研究中未测出。其主要成分依次为亚油酸、棕榈酸、油酸、硬脂酸，其中亚油酸含量最高（种子81.74%，果肉66.97%），致使不饱和脂肪酸含量较高（种子89.89%，果肉81.49%），营养价值较高，具有高档食用油开发前景。（3）未经低温处理的不同分布区的山桐子种子在15/25℃(15℃12个小时，25℃12个小时)发芽温度下发芽率存在显著差异，变幅为10.72%～65.13%；发芽率最高为广元，最低为临安。变化规律与纬度没有明显的相关性，从总体上来看，除临安分布区外，表现为边缘分布区的发芽率大于中心分布区。在低温处理一定时间后，不同分布区的种子发芽率均有不同程度的提高，其中发芽率最高的为南昌，条件为低温10℃20d，高达85.75%；发芽率最低的为临安，条件为10℃60d，低至22.96%。不同分布区最合适的低温处理温度和低温处理时间不同，且随着纬度的增加，山桐子解除休眠的适合温度呈增加趋势。总体而言，从处理温度来看，大部分10℃的低温解除其休眠效果较好，说明山桐子种子休眠深度较浅，只需要少量的低温量即可解除其休眠。从处理时间来看，大部分处理20d解除其休眠效果较好，处理温度的增高和处理时间的继续增加反而抑制了部分种子的萌发。（4）未经低温处理的不同分布区的山桐子种子在15/25℃发芽温度下发芽势存在明显差异，变幅为5.67%～35.46%；发芽势最高为成都，高达35.46%，最低为重庆，低至5.67%。在低温处理一定时间后，不同分布区的种子发芽势均有不同程度的提高，不同分布区最合适的低温处理温度和低温处理时间不同，总体而言，从处理温度来看，10℃的低温提高发芽势的效果较好，从处理时间来看，处理20d和40d提高其发芽势效果较好。（5）未经低温处理的不同分布区的山桐子种子在15/25℃发芽温度下发芽速度存在明显差异，变幅为13.48～20.56d；发芽时间最长为张家界20.56d，最短为成都13.18d。不同分布区山桐子种子在不同低温处理不同时间的最长发芽时间和最短发芽时间有较大差异，从总体来看，不同分布区山桐子经过不同时间不同温度低温处理下的种子发芽时间最长的温度均为15℃，说明山桐子种子在较长时间较高温度的条件下，种子发芽速度与未处理相比，反而降低。（6）不同分布区山桐子发芽指标与种实品质有一定的相关关系。发芽率与种子千粒重、种子含油率、果肉含油率、果实含油率呈显著正相关，发芽势与种子含油率、果肉含油率、果实含油率、种子碘值呈显著正相关，与种子十六烷值呈显著负相关。发芽时间与果实品质关系不大。（7）采用改良CTAB法，通过调整裂解液浓度比例，控制裂解时间和添加PVP40和β-巯基乙醇等方法，摸索出了适合山桐子叶片基因组DNA的提取方法，可满足用于ISSR遗传分析。筛选出的22条引物，其中第8条IS44序列（GAC）5，说明山桐子的基因组中存在有寡聚三核苷酸重复序列。用选出的10个引物对12个分布区共110个DNA样品进行PCR扩增，共扩增出92条重复性高、清晰的条带，扩增片段分子量为200～2500bp之间，平均每个引物扩增位点为9.2个，各引物扩增出的条带数为6～11条，其中多态性位点85个，多态性比率为87.5%～100%，平均多态性比率为92.91%。（8）不同分布区山桐子的遗传多样性存在一定差异。总体来看，位于山桐子分布中心区的广元、重庆、张家界、成都、宜昌有较高的遗传多样性，而位于分布区边缘的赣州、遵义、洛阳、九江、贵阳、临安南昌种群遗传多样性偏低，但没有随纬度的变化而呈现出有规律性的变化。山桐子分布区间与分布区内都产生了显著的遗传分化（p<0.001），山桐子的遗传多样性主要在分布区内，占变异成分的78.01%，只有21.99%的变异存在于不同种群间。说明山桐子的变异主要存在分布区内，分布区间的遗传分化较小。（9）进行UPGMA聚类分析，当相似系数为0.868时，可聚为4类，九江为一类，赣州为一类，南昌为一类，其余9个分布区为一类。不同分布区之间的遗传一致度在0.7236～0.9806之间，说明不同分布区山桐子群体间的相似程度较高。其中重庆和张家界两个分布区的遗传距离最小为0.0196，南昌和九江两个分布区的遗传距离最大为0.3235。山桐子分布区间的遗传距离并没随纬度变化而呈现明显的规律变化。将遗传距离与地理距离做相关性分析，显著水平p＞0.05,说明山桐子不同分布区间遗传距离与地理距离没有相关性。更多还原

【Abstract】 This paper mainly analyses the quality difference, seed dormancydiversity, genetic variability and correlation with ecology and geography of Idesiapolycarpa seeds located in Ganzhou(Jiangxi province), Guiyang and Zunyi (GuizhouProvince),Jiujiang and Nanchang (Jiangxi Province), Zhangjiajie (Hunan Province),Lin’an (Zhejiang Province), Yichang (Hubei Province), Chengdu and Guangyuan(Sichuan Province), Chongqing and Luoyang (Henan Province)12provenances anddraws a Idesia polycarpa geographic population variation. Main conclusions are asfollows:（1）Oil content of Idesia polycarpa pulp of12provenances was28.38%~48.35%, average37.62%, seed oil content12.6%~28.17%, average21.17%,average fruit (seeds and pulp) oil content was31.36%, and the Idesia polycarpa seediodine value between112.6~139.7mg·g-1, saponification value between191.2~194.6mg·g-1, cetane value between43.2~49.3mg·g-1, the iodine value of the pulpbetween108.2~137.1mg·g-1, saponification value between196.2~205.6mg·g-1,cetane value between43.3~55.0mg·g-1.According to the evaluation standard ofbiodiesel products, Idesia polycarpa is a kind of high quality raw materials ofbiodiesel. Guiyang area can be drawn as the most appropriate raw materials area forthe biomass diesel oil and can be used as an Idesia polycarpa priority to thedevelopment of biomass energy forestry region, following by Luoyang and Ganzhou.The fruit quality differences of Idesia polycarpa in different areas have a certainrelationship with the climate conditions of each area, the main factors being effectiverainfall and shining hours.（2）The Idesia polycarpa fruit oil has as high as12kinds of fatty acids (seed is10,12kinds of pulp). This experiment measured9-hydroxy pelargonic acid, twelveacids, heptadecanoic acid which predecessors did not find. The main ingredients arelinoleic acid, palmitic acid, oleic acid, stearic acid. The linoleic acid content is thehighest (seed81.74%,66.97%) pulp, the unsaturated fatty acid content is higher(seed89.89%,81.49%pulp), having a bright future for edible oil development.（3）Idesia polycarpa seeds without low temperature treatment in the15/25℃(12hours25℃,15℃for12hours) exist obvious differences. The variance testshows significant differences (P <0.05), the range of10.72%to10.72%; the highest germination rate of Guangyuan, as high as65.13%, the lowest for Lin’an, as low as10.72%. In the order: Guangyuan (65.13%)> Nanchang (61.72%)> Jiujiang (57.93%)> Luoyang (44.92%)> Chengdu (42.83%)> Zunyi (36.61%)> Ganzhou (33.00%)> Yichang (32.54%)> Zhangjiajie (26.00%)> Guiyang (23.00%)> Chongqing(16.00%)> Lin’an (10.72%). Change rules have no obvious correlation with latitude,as a whole, in addition to Lin’an distribution zone, the germination rate at the edge ofthe area is greater than the center area. Under low temperature treatment after acertain time, seed germination rate of different areas have different degree of increase,in which the seeds of Zunyi area under the condition of5℃to60days of treatment,germination rate increased to72.02%from36.61%when untreated, the maximumbeing35.41%and increased to96.72%. Under the same process, Idesia polycarpa ofdifferent areas have significant differences in seed germination rate. The variance testshows that under the same process, different significant differences were observed ingermination rate (P <0.05). In a word, the Idesia polycarpa seeds treated by lowtemperature are higher than that without low temperature treatment in seedgermination rate. In Lin’an,15℃treatments under the condition of60days, seedgermination rate is zero, which may be due to seed at high temperatures for a longtime which in turn, inhibit the seed germination.（4）The seed germination potential of different Idesia polycarpa seeds withoutlow temperature treatment exists obvious differences, under the15/25℃germinatingtemperature, the variance test showing significant differences (P <0.05), the range of5.67%to5.67%; Germinability of Chengdu is the highest, up to35.46%, the lowestfor Chongqing, as low as5.67%. After a certain time in low temperature processing,seed germination potential of different areas all have varying degrees of increase. Indifferent distribution areas, the most suitable low temperature processing tempratureand low temperature processing time are different. In general, from the point ofprocessing time, processing of20and40days to improve the germination potentialeffect is better.（5）The germination speed of different Idesia polycarpa seeds without lowtemperature treatment germinate in the15/25℃temperature exists obviousdifference, the variance test showing significant difference (P <0.05), the range of13.48to13.48days. Overall, the longest seed germination time of Idesia polycarpafrom different area at different time under different temperature and low temperatureprocessing of is15℃, showing that the seed germination rate of Idesia polycarpa seeds under the condition of high temperature and a long time, compared with theuntreated, reduces instead.（6）The germination index and the quality of Idesia polycarpa in differentareas have certain correlations. The correlations between seed germination rate andTKW, the seed oil content, pulp oil content, the oil content of the fruit arepositive.The germination tendency is negatively associated with the seed oil content,pulp oil content, the oil content of the fruit, seed iodine value. Germination time islittle associated with fruit quality.（7）The results of DNA extraction and primers screening show that owing tothe high Idesia polycarpa polyphenols content in leaf, samples are prone to Browning,difficult to extract DNA. This research adopts the improved CTAB method, byadjusting the pyrolysis liquid concentration ratio, to control the cracking time, addPVP40and beta mercaptoethanol, grope for the table leaf genomic DNA extractionmethod, can satisfy the ISSR genetic analysis. This study selected22primers inarticle8GACGACGACGACGAC IS44sequences, that Idesia polycarpa isoligomeric trinucleotide repeat sequences in the genome. Using selected10primers of12areal PCR amplification, a total of110DNA samples were amplified from92highrepeatability, clear bands of fragments amplified molecular weight between200~2500bp, the average each primer amplification loci was9.2, the primer amplificationof stripe number for polymorphism loci of6and11,85, the average polymorphismrate was92.91%.（8）There exist certain differences between12Idesia polycarpa species. In aword, however, the Idesia polycarpa of the distribution centers, Zhangjiajie, Chengdu,Guangyuan, Chongqing, and Yichang had a higher genetic diversity, and Ganzhou,Zunyi, Luoyang, Lin’an Nanchang, Jiujiang, Guiyang which are located in the edge ofthe area had low genetic diversity of population. But there is no change with latitudeand presenting the regularity of change. Idesia polycarpa distribution range anddistribution area had significant genetic differentiations (p <0.001), the geneticdiversity of Idesia polycarpa in the main distribution area, accounting for78.01%ofthe variation of composition, only21.99%of the variation existed between differentpopulations. All this show that Idesia polycarpa variations mainly exist in distributionarea, distribution range of genetic differentiation being smaller.（9）In doing UPGMA cluster analysis, when the similarity number was0.868,the areas can be classified into4groups, a class for Jiujiang, a class for Ganzhou, a class for Nanchang, the remaining9areas together for a class. The geneticconsistency between different areas is0.7236to0.9806, which shows high degree ofsimilarity between different Idesia polycarpa of distribution areas. The minimumgenetic distances of Chongqing and Zhangjiajie is0.0196, the maximum geneticdistance of Nanchang and Jiujiang0.3235. Distribution intervals of the geneticdistance don’t change regularly with latitude. After doing the correlation analysis ofgenetic distance and geographic distance, significant level being (p>0.05), thegenetic distance of desia polycarpa in different distribution were not linked togeographic distance.更多还原