何首乌野生种质资源地理变异与种源初步选择
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摘要
何首乌(Polygonum multilorum Thunb.)为蓼科的多年生缠绕草质藤本植物,其块根及藤均为名贵的大宗中药材,在保健和化工等方面也有广阔的开发前景。
本文对全国10 个省份何首乌野生种源的遗传差异和生理、生长及质量性状差异进行研究,取得的主要研究结果如下:
1.通过主成分和聚类分析方法,依据叶性状,将31 个何首乌野生种源划分为三大类。
2.通过改进CTAB 法提取出高质量的DNA,首次建立了RAPD 标记在该物种上的良好反应体系。
3.从300 对引物中筛选出40 对稳定且重复性好的引物进行扩增,发现野生何首乌种源具有较高的多态性(69.5%);所检测到的228 条多态位点的平均有效等位基因数目(Effective number of alleles)为1.4556,平均基因多样度(Nei's gene diversity)为0.2781,平均信息指数(Shannon's Information index)为0.4329,研究显示了何首乌野生种源具有较丰富的遗传多样性;聚类分析将31 个何首乌野生种源划分为四大类,与叶性状的地理变异基本相吻合,表明各种源间的亲缘关系与地理分布相符。
4.扦插试验结果表明31 个种源的扦插成活率有显著差异;茎段组织培养研究表明何首乌芽的芽增殖培养配方以MS+6-BA1.0mg/L+NAA0.05mg/L 和MS+ 6-BA0.5mg/L+NAA0.02 mg/L培养基较好,生根诱导以1/2MS+6-BA1.0mg/L +NAA0.1mg/L或1/2MS+ 6-BA0.5mg/L +NAA0.05mg/L 培养基较好
5.研究表明: 10 个何首乌种源在生长、生理及质量各性状方面均存在显著或极显著差异,其中:(1)普定、环江和南京种源的总藤长生长迅速,高州、泰山和南京种源的块茎生长较快,而高州、环江和恩施种源的总生物量较高;(2)北方种源的光合速率比南方种源略高,但在单株光合产率上,环江、高州和恩施等南方种源较高,这与总生物量的表现一致;此外南方种源的PSⅡ光化学效率比北方种源高;各种源叶绿素含量、可溶性蛋白质含量、硝酸还原酶活性和过氧化物酶活性在各月份的变化差异显著;(3)何首乌各种源在块根、藤和叶片中的6 种金属元素含量变化差异显著;(4)泰山、嵩县和峨眉种源的总蒽醌含量较高,但高州和泰山种源的结合型蒽醌有效经济产量高于其它种源。
6.通过性状间相关分析,选出11 个性状指标作为何首乌种源选择的指标。
7.通过主成分分析和因子得分模型分析,初步筛选出2 个适合在江苏省推广的优良何首乌种源。
Polygonum multiflorum is perennial herbaceous vine and it’s tuber and vine are used as traditional medicine. Polygonum multiflorum is poplar in many aspects, such as health protection and chemical engineering.
The main objective of this thesis is to study difference in genetic, physiology, growth, and chemical compounds among Polygonum multiflorum provenances. The main results are as following:
1. Base on the principle component analysis method and the cluster analysis method, 31 wild provenances of Polygonum multiflorum were divided into three classes by leaf characters.
2. CTAB method was refined to extract high quality DNA, and good reaction system of RAPD marker was built for the species for the first time.
3. 40 pairs primes of high stability and good repeat ability were selected from 300 pairs to amplify. The results showed that wild Polygonum multiflorum provenances had high polymorphic locus percentage (69.5%). The average effective number of alleles in 228 polymorphic loci detected in the test was 1.4556, the average Nei's gene diversity was 0.2781, and the average Shannon's information index was 0.4329. The study showed that wild Polygonum multiflorum provenances had abundant gene diversity. According to the cluster analysis method, 31 wild provenances of Polygonum multiflorum were divided into four classes, generally according with geographic variation of leaf character. The results showed that the relative relationship among different provenances was corresponded with geographical distribution.
4. Vine cutting showed that there were significantly differences in the cutting survival rate among 31 provenances. The tissue culture showed that the proliferation culture prescription used for Polygonum multiflorum bud culture was MS+6-BA1.0mg/L+NAA0.05mg/L and MS+6-BA0.5 mg/L + NAA0.02 mg/L, and the culture medium of rooting was 1/2MS+6-BA1.0mg/L+ NAA0.1 mg/L or 1/2MS+6-BA0.5mg/L+ NAA 0.05mg/L.
5. The study showed that there were significant differences in the growth characters, physiology characters, and quality characters among 10 Polygonum multiflorum provenances. (1) Of all testing materials, provenances in PuDing, HuanJiang and NanJing had fast vine growth; provenances in GaoZhou, TaiShan, and NanJing had best underground tuber growth, and provenances in GaoZhou, HuanJiang and EnShi had best biomass. (2) Net photosynthetic rate of north provenances was a litter higher than that of south provenances. The photochemical efficiency of PSII of south provenances was higher than that of north provenances. There were significant variation on chlorophyll content, solubility proteins content, and nitrate reductaes activity and peroxide enzyme activity among provenances. (3) Among different provenances of Polygonum multiflorum, there were significant variation on
content of 6 microelements in the tuber, vine and leaf. (4) Provenances of in TaiShan, SongXian and EMei had higher the total anthraquinones content than others. However, provenances in GaoZhou and TaiShan had higher the effective economic yield of anthraquinones than others.
6. Relative analysis among characters showed there were significantly negative relativity between 10 characters and the tuber, and 11 different goal character targets were selected out as the important target.
7. By principle analysis and factor analysis, two good provenances were initially selected to be adapted to planting in Jiangsu province.
本文对全国10 个省份何首乌野生种源的遗传差异和生理、生长及质量性状差异进行研究,取得的主要研究结果如下:
1.通过主成分和聚类分析方法,依据叶性状,将31 个何首乌野生种源划分为三大类。
2.通过改进CTAB 法提取出高质量的DNA,首次建立了RAPD 标记在该物种上的良好反应体系。
3.从300 对引物中筛选出40 对稳定且重复性好的引物进行扩增,发现野生何首乌种源具有较高的多态性(69.5%);所检测到的228 条多态位点的平均有效等位基因数目(Effective number of alleles)为1.4556,平均基因多样度(Nei's gene diversity)为0.2781,平均信息指数(Shannon's Information index)为0.4329,研究显示了何首乌野生种源具有较丰富的遗传多样性;聚类分析将31 个何首乌野生种源划分为四大类,与叶性状的地理变异基本相吻合,表明各种源间的亲缘关系与地理分布相符。
4.扦插试验结果表明31 个种源的扦插成活率有显著差异;茎段组织培养研究表明何首乌芽的芽增殖培养配方以MS+6-BA1.0mg/L+NAA0.05mg/L 和MS+ 6-BA0.5mg/L+NAA0.02 mg/L培养基较好,生根诱导以1/2MS+6-BA1.0mg/L +NAA0.1mg/L或1/2MS+ 6-BA0.5mg/L +NAA0.05mg/L 培养基较好
5.研究表明: 10 个何首乌种源在生长、生理及质量各性状方面均存在显著或极显著差异,其中:(1)普定、环江和南京种源的总藤长生长迅速,高州、泰山和南京种源的块茎生长较快,而高州、环江和恩施种源的总生物量较高;(2)北方种源的光合速率比南方种源略高,但在单株光合产率上,环江、高州和恩施等南方种源较高,这与总生物量的表现一致;此外南方种源的PSⅡ光化学效率比北方种源高;各种源叶绿素含量、可溶性蛋白质含量、硝酸还原酶活性和过氧化物酶活性在各月份的变化差异显著;(3)何首乌各种源在块根、藤和叶片中的6 种金属元素含量变化差异显著;(4)泰山、嵩县和峨眉种源的总蒽醌含量较高,但高州和泰山种源的结合型蒽醌有效经济产量高于其它种源。
6.通过性状间相关分析,选出11 个性状指标作为何首乌种源选择的指标。
7.通过主成分分析和因子得分模型分析,初步筛选出2 个适合在江苏省推广的优良何首乌种源。
Polygonum multiflorum is perennial herbaceous vine and it’s tuber and vine are used as traditional medicine. Polygonum multiflorum is poplar in many aspects, such as health protection and chemical engineering.
The main objective of this thesis is to study difference in genetic, physiology, growth, and chemical compounds among Polygonum multiflorum provenances. The main results are as following:
1. Base on the principle component analysis method and the cluster analysis method, 31 wild provenances of Polygonum multiflorum were divided into three classes by leaf characters.
2. CTAB method was refined to extract high quality DNA, and good reaction system of RAPD marker was built for the species for the first time.
3. 40 pairs primes of high stability and good repeat ability were selected from 300 pairs to amplify. The results showed that wild Polygonum multiflorum provenances had high polymorphic locus percentage (69.5%). The average effective number of alleles in 228 polymorphic loci detected in the test was 1.4556, the average Nei's gene diversity was 0.2781, and the average Shannon's information index was 0.4329. The study showed that wild Polygonum multiflorum provenances had abundant gene diversity. According to the cluster analysis method, 31 wild provenances of Polygonum multiflorum were divided into four classes, generally according with geographic variation of leaf character. The results showed that the relative relationship among different provenances was corresponded with geographical distribution.
4. Vine cutting showed that there were significantly differences in the cutting survival rate among 31 provenances. The tissue culture showed that the proliferation culture prescription used for Polygonum multiflorum bud culture was MS+6-BA1.0mg/L+NAA0.05mg/L and MS+6-BA0.5 mg/L + NAA0.02 mg/L, and the culture medium of rooting was 1/2MS+6-BA1.0mg/L+ NAA0.1 mg/L or 1/2MS+6-BA0.5mg/L+ NAA 0.05mg/L.
5. The study showed that there were significant differences in the growth characters, physiology characters, and quality characters among 10 Polygonum multiflorum provenances. (1) Of all testing materials, provenances in PuDing, HuanJiang and NanJing had fast vine growth; provenances in GaoZhou, TaiShan, and NanJing had best underground tuber growth, and provenances in GaoZhou, HuanJiang and EnShi had best biomass. (2) Net photosynthetic rate of north provenances was a litter higher than that of south provenances. The photochemical efficiency of PSII of south provenances was higher than that of north provenances. There were significant variation on chlorophyll content, solubility proteins content, and nitrate reductaes activity and peroxide enzyme activity among provenances. (3) Among different provenances of Polygonum multiflorum, there were significant variation on
content of 6 microelements in the tuber, vine and leaf. (4) Provenances of in TaiShan, SongXian and EMei had higher the total anthraquinones content than others. However, provenances in GaoZhou and TaiShan had higher the effective economic yield of anthraquinones than others.
6. Relative analysis among characters showed there were significantly negative relativity between 10 characters and the tuber, and 11 different goal character targets were selected out as the important target.
7. By principle analysis and factor analysis, two good provenances were initially selected to be adapted to planting in Jiangsu province.
引文
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