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绿竹遗传多样性与优良地理种源选择的研究

Genetic Diversity and Selection of Super Geographical Provenances of Dendrocalamopsis Oldhami

【作者】 郑蓉

【导师】 方伟;

【作者基本信息】 北京林业大学 , 森林培育, 2008, 博士

【摘要】 绿竹(Dendrocalamopsis oldhami(Munro) Keng f.)是我国南方重要的经济生态型丛生竹之一,在中亚热带以南地区有广泛分布,各地理区域表现出植物形态、生理生化、笋品质和适应性等差异,针对目前绿竹引种的局域性和盲目性等问题,为明确其遗传基础及生态适应范围,本研究在绿竹资源的表型、笋品质、DNA分子标记分析基础上,收集国内主要产区的绿竹地理种源进行造林试验,从造林成活、生长物候、产笋性能、越冬生理、生态适应性等方面系统研究,综合评价出优良地理种源/单丛:1、对12个产地绿竹的表型性状与笋品质分析表明,绿竹资源具有较为丰富的形态性状和品质性状的多样性。12个主要形态学性状的变异系数为5.5-27.2%,表明不同产地绿竹、不同性状之间存在着差异,其中枝下高、叶片长、叶片宽、胸径、竹高等变异系数大于10%;竹高、枝下高分别与叶长、叶宽的相关性达显著极显著水平,叶长、节数与竹高也具有显著相关性,这些性状也是衡量绿竹生产性能和适应性的重要形态指标。聚类图表明各地绿竹生长性状、笋养分指标呈现出一定地域性差异,但不明显,典型相关分析进一步反映笋营养成分与土壤养分指标有关联性。2、以各地绿竹21个样品及其2个近缘种为DNA提取材料,初步建立了AFLP体系。选用的20对选择性引物组合通过该体系对23个样品扩增,银染后得到较清晰的条带,并明显区分出2个近缘种,但绿竹样品之间扩增产物尚未检测出差异性条带。3、对GenBank中绿竹(Bambusa oldhamii)EST数据库中的序列分析设计出16对引物。选择5对引物的PCR产物完成测序,含有预测的SSR位点,2对引物在24个样品中扩增产物没有位点的多态性,3对引物扩增的位点在广州绿竹、乌脚绿竹(Dendrocalamopsis edulis)和苦绿竹(D. basihirsuta)样品中有多态性。4、自然变温条件下,绿竹地理种源的保护酶活性、可溶性蛋白、可溶性糖含量、叶绿素含量及叶绿素荧光参数比较分析得,抗寒性从强到弱依次为:温州绿竹>福安绿竹>尤溪绿竹>漳州绿竹,隶属度值分析也表明温州绿竹抗寒性稍强,证实了绿竹越冬能力与其原产地环境是相适应的。5、绿竹地理种源试验各测定指标与遗传效应分析表明,2个试验点各地理种源的造林成活率差异较大,但出笋物候期无显著差异。9个供试种源在漳州均能正常生长,较好地适应引种地生态气候条件;幼林期与成林期的高径生长种源间差异显著;笋产量的稳定程度大小依次为D2、D3、D1、D5、D7、D9、D6、D4、D8,随着竹林郁闭成林后,各种源的笋产量逐步趋向稳定;各种源年平均叶面积指数、叶绿素含量、光合速率、呼吸速率等指标也均存在一定的差异,但不同指标的种源排序各异。应用五分评定法对地理种源的生态适应性、产笋性能的评定结果与运用主成分分析9个主要产量性状的结果基本一致,筛选出适合在南亚热带栽培的优良绿竹种源与单丛,入选地理种源有D1、D2、D9、D6、D7、D5,入选单丛有D2-6、D7-9、D3-10、D5-11等,主要性状的现实增益在6.2%-34.6%之间。

【Abstract】 Dendrocalamopsis oldhami is one of the important economical and ecotypic species of clumpy bamboo in south China, which is widely distributed in the southern regions of mid-subtropics, and there are different cultivation qualities in all planting regions. In order to settle the introduction problem and know heredity foundation and the range of ecological adaptation, the traits on phenotypic, qualities of shoot and DNA genetic marker were analyzed in all resource, geographic provenance that distributed in the main production region of China were collected for forestation, and excellent provenances were evaluated by growth period, production performance, ecological adaptability, etc.1. Phenotypic traits and qualities of shoot were studied among 12 provenances. The result showed that diversity of morphological and economic traits of D. oldhami are abundant. 12 main coefficient of variation(CV) of morphological traits are range 5.5% to 27.2%, the CV of live branch height, length and width of leaf, diameter of breast height, tree height are more than 10%. The significance level of correlation are different between botanic phenotypic traits of D. oldhami. The correlation between tree height, live branch height and length of leaf, width of leaf were significant and great significant, and significant between length, numbers of pieces and tree height, these traits are important morphological indicators about yield capability of D. oldhami. According to clustering analysis, the growth traits of bamboo and qualities of shoot are a few difference from different original, and there is certain correlation depended on soil and climate environment.2. An AFLP analysis system is established for genome DNA of 21 specimens from geographic provenances of D. oldhami and 2 species of Dendrocalamopsis. The 23 samples can be amplified by this system with twenty couples of selective primers. Obvious silver-staining bands can be found by electrophoresis separation with 6% metamorphic polyacrylamide gel in PCR production, and 2 species of Dendrocalamopsis can be distinguished easily. Whereas, there are a few polymorphic band among the amplification production in the samples.3. According to the Expressed Sequence Tags(EST) database about Bambusa oldhamii in GenBank, sixteen couples of primers were designed. The amplified production of PCR by 5 couples of primer were complete mapped genome, there were simple sequence repeats(SSRs) markers. Twenty-four specimens of genome DNA were tested by 5 couples of primer, no polymorphic bands be found through the amplified productions by 2 couples of primer, and none of 21 samples except samples from Guangzhou provenance, samples of Dendrocalamopsis edulis and D.basihirsuta by 3 couples of primer.4. Under overwintering condition, the properties were analyzed including the activity of protective enzymes and the content of soluble protein, soluble sugar, chlorophyll content and chlorophyll fluorescence parameters from different provenance. The range of cold resistance of provenance are as follows: Wenzhou> Fuan> Youxi >Zhangzhou, and the membership degree value from Wenzhou is much higher than from others, that is the best explanation of higher cold resistance from Wenzhou than from others.5. The results of evaluation on traits and genetic parameters shows that the survival rates of planting are different among geographic provenances in 2 test sites, the period of shoot growth are similar. 9 provenances accomplished normal growth, and easily adapted to the ecologic climate conditions in Zhangzhou site. The height and diameter growth were significant difference between the youth and adult forest. The stability of shoot products were analyzed, the value from high to low as follows:D2, D3, D1, D5, D7, D9, D6, D4, D8. Overall, the shoot products tend to be stable after the crown closed. The result of study on traits of leaf shows that there are difference of leaf area index, chlorophyll content, photosynthesis rate, and respiration rate between different provenance, and the difference of orders of different index. The same results were brought not only by five-grade evaluation on ecological adaptability and ability of shoot yield, but also by principal component analysis on main traits. Superior provenances adapting to the south of sub-tropic were selected, and the practical plus range from 6.2% to 34.6% in average.

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