118份甘蔗种质资源遗传多样性的AFLP分析

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昝逢刚, 吴才文, 陈学宽, 赵培方, 赵俊, 刘家勇. 118份甘蔗种质资源遗传多样性的AFLP分析. 作物学报, 2014, 40(10): 1877-1883[ZAN Feng-Gang, WU Cai-Wen, CHEN Xue-Kuan, ZHAO Pei-Fang, ZHAO Jun, LIU Jia-Yong. Genetic Diversity of 118 Sugarcane Germplasm Using AFLP Markers. Acta Agronomica Sinica, 2014, 40(10): 1877-1883] 复制到剪切板

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118份甘蔗种质资源遗传多样性的AFLP分析

昝逢刚, 吴才文, 陈学宽, 赵培方, 赵俊, 刘家勇^*

云南省农业科学院甘蔗研究所 / 云南省甘蔗遗传改良重点实验室, 云南开远661699

^* 通讯作者(Corresponding author): 刘家勇, E-mail:lljjyy1976@163.com

收稿日期:2014-07-06

基金:本研究由国家现代农业产业技术体系建设专项(CARS-20-1-1), 云南省应用基础研究面上项目(2010CD003)和云南省重点新产品开发计划项目(2012BB014)资助;

摘要

采用AFLP标记技术, 对来自中国、澳大利亚、美国、菲律宾、巴西和法国的118份甘蔗种质进行遗传多样性分析。采用10对引物组合共扩增出1310条谱带, 其中多态性条带为1195条, 多态性条带比率为91.2%。118份种质间的相似性系数在0.583~0.929之间, 平均为0.750, 多态信息量为0.2332, 每个位点的有效等位基因数为1.3789。结果显示在相似性系数0.69处切割时, 可划分为2个类群, 第I类群包括粤糖00-236、云蔗04-622和SP80-1816; 第II类群有115份种质, 在相似性系数0.74处切割时, 可将第II类群划分为5个亚群。各国甘蔗种质亲缘关系较近, 其中美国种质遗传多样性相对较丰富。

关键词: 甘蔗; 种质; AFLP; 遗传多样性

Genetic Diversity of 118 Sugarcane Germplasm Using AFLP Markers

ZAN Feng-Gang, WU Cai-Wen, CHEN Xue-Kuan, ZHAO Pei-Fang, ZHAO Jun, LIU Jia-Yong^*

Sugarcane Research Institute, Yunnan Academy of Agricultural Sciences / Yunnan Province Key Laboratory of Sugarcane Genetic Improvement, Kaiyuan 616699, China

Fund:

Abstract

AFLP markers were used to assess the genetic diversity of 118 sugarcane germplasm from China, Australia, USA, Philippines, Brazil, and France. A total of 1310 bands were amplified with ten pairs of primers, 1195 of which were polymorphic bands, the percentage of polymorphic loci was 91.22%. The genetic similarity coefficient among 118 sugarcane germplasm ranged from 0.583 to 0.929 with an average of 0.750, the polymorphism information content was 0.2332, and the effective number of alleles per loci was 1.3789. The results showed 118 germplasm were divided into two groups at 0.69 of the similarity coefficient. The group I consisted of YT00-236, YZ04-622, and SP80-1816, which shared lower genetic similarity with the rest of 115 germplasm in another group. The 115 germplasm were divided into five sub-groups at 0.74 of the similarity coefficient. The sugarcane germplasm shared high genetic similarity among countries, and there existed much higher genetic diversity within germplasm introduced from USA.

Keyword: Sugarcane; Germplasm; AFLP; Genetic diversity

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甘蔗蔗糖产量约占我国食糖总量的92%, 具有遗传多样性的甘蔗种质资源是甘蔗杂交育种的基础。目前, 甘蔗种质直接用于杂交利用较为普遍, 且作用巨大, 国外种质资源在我国杂交育种中也具有更重要的作用, 我国利用引进种质Co419、CP49-50、NCo310^{[ 1]}和CP72-1210^{[ 2]}, 分别育成了24、26、27和12个优良品种。甘蔗优良品种选育的主要途径是杂交育种, 组合选配则是选育优良品种成败的关键, 组合选配得好, 其后代就会出现较多的优良变异, 就有可能选育出新的优良品种, 因此研究不同种质的遗传关系对组合选配具有重大意义。表型性状极易受环境因素影响而变化, 仅依靠表型性状难以准确反映不同材料间的遗传差别和亲缘关系^{[ 3, 4]}。品种系谱虽然在一定程度上能反映品种之间的亲缘关系, 但由于自交、串粉等因素, 存在一定的局限性^{[ 5, 6, 7]}, 基于分子水平的亲缘关系较基于系谱的亲缘关系具有更高的可靠性^{[ 8, 9]}。

AFLP标记技术综合了RFLP和RAPD的优点, 既具有RFLP的稳定性和PCR反应快速、灵敏的特点, 又具有RAPD的方便性, 同时还克服了RFLP和RAPD的缺点^{[ 10]}。由于AFLP标记多态性强, 结果稳定, 重复性好, 是一种理想、有效的遗传标记, 近年被广泛用于农作物的遗传多样性、遗传图谱、基因定位、指纹图谱等研究^{[ 3, 4, 11, 12]}。李鸣等^{[ 13]}采用AFLP技术, 用5对引物扩增在生产上广泛推广的2个甘蔗品种, AFLP指纹图谱分析区分率达100%。庄南生等^{[ 8]}采用AFLP分子标记技术分析54份甘蔗种质(14份祖亲种、40份栽培品种或品系)的遗传基础, 利用筛选出的4对多态性较强的引物组合, 构建了甘蔗54份种质的AFLP指纹图谱。刘新龙等^{[ 14]}采用SSR和AFLP标记技术, 对构建的2个群体进行PCR扩增和分子遗传连锁分析,构建了甘蔗分子遗传连锁图谱。本文基于AFLP分子标记, 分析国内外118份甘蔗种质的遗传多样性, 旨在明确种质间的亲缘关系, 为甘蔗种质的杂交利用和种质创新提供参考依据。

1 材料与方法

1.1 试验材料

118份种质采自云南省农业科学院甘蔗研究所的国家甘蔗种质资源圃, 其中中国33份, 澳大利亚19份, 美国16份, 菲律宾22份, 巴西13份, 法国15份(表1)。

表1 试验种质名称及来源 Table 1 Name and origin of germplasm used in the experiment

(续表1) 试验种质名称及来源 Table 1(Continued) Name and origin of germplasm used in the experiment

序号 No.	名称 Name	来源 Origin	序号 No.	名称 Name	来源 Origin
18	云蔗04-488 Yunzhe 04-488	中国 China	77	FR99-378	法国 France
19	云蔗04-621 Yunzhe 04-621	中国 China	78	Q107	澳大利亚 Australia
20	云蔗04-622 Yunzhe 04-622	中国 China	79	Q120	澳大利亚 Australia
21	云蔗04-649 Yunzhe 04-649	中国 China	80	Q127	澳大利亚 Australia
22	云蔗04-724 Yunzhe 04-724	中国 China	81	Q158	澳大利亚 Australia
23	云蔗05-181 Yunzhe 05-181	中国 China	82	Q162	澳大利亚 Australia
24	云蔗05-197 Yunzhe 05-197	中国 China	83	Q171	澳大利亚 Australia
25	云蔗05-66 Yunzhe 05-66	中国 China	84	Q174	澳大利亚 Australia
26	云蔗06-102 Yunzhe 06-102	中国 China	85	Q182	澳大利亚 Australia
27	云蔗06-267 Yunzhe 06-267	中国 China	86	Q188	澳大利亚 Australia
28	云蔗06-407 Yunzhe 06-407	中国 China	87	Q192	澳大利亚 Australia
29	云蔗89-7 Yunzhe 89-7	中国 China	88	Q195	澳大利亚 Australia
30	云蔗94-343Yunzhe 94-343	中国 China	89	Q196	澳大利亚 Australia
31	云蔗94-375 Yunzhe 94-375	中国 China	90	Q197	澳大利亚 Australia
32	云蔗99-596 Yunzhe 99-596	中国 China	91	Q198	澳大利亚 Australia
33	云蔗99-91 Yunzhe 99-91	中国 China	92	Q199	澳大利亚 Australia
34	CP33-310	美国 USA	93	Q202	澳大利亚 Australia
35	CP67-412	美国 USA	94	Q207	澳大利亚 Australia
36	CP81-1302	美国 USA	95	Q209	澳大利亚 Australia
37	CP81-1405	美国 USA	96	Q88	澳大利亚 Australia
38	CP85-1382	美国 USA	97	VMC73-229	菲律宾 Philippines
39	CP86-1633	美国 USA	98	VMC81-202	菲律宾 Philippines
40	CP86-1644	美国 USA	99	VMC84-549	菲律宾 Philippines
41	CP88-1546	美国 USA	100	VMC87-599	菲律宾 Philippines
42	CP88-1834	美国 USA	101	VMC87-095	菲律宾 Philippines
43	CP89-2143	美国 USA	102	VMC90-239	菲律宾 Philippines
44	CP89-2376	美国 USA	103	VMC93-331	菲律宾 Philippines
45	CP89-2377	美国 USA	104	VMC93-341	菲律宾 Philippines
46	CP92-1641	美国 USA	105	VMC94-050	菲律宾 Philippines
47	CP94-1100	美国 USA	106	VMC94-266	菲律宾 Philippines
48	HOCP96-540	美国 USA	107	VMC95-018	菲律宾 Philippines
49	TCP83-3196	美国 USA	108	VMC95-009	菲律宾 Philippines
50	RB73-2727	巴西 Brazil	109	VMC95-105	菲律宾 Philippines
51	RB73-9735	巴西 Brazil	110	VMC95-262	菲律宾 Philippines
52	RB75-11	巴西 Brazil	111	VMC95-088	菲律宾 Philippines
53	RB83-5054	巴西 Brazil	112	VMC96-134	菲律宾 Philippines
54	RB83-5089	巴西 Brazil	113	VMC96-169	菲律宾 Philippines
55	RB85-5113	巴西 Brazil	114	VMC96-055	菲律宾 Philippines
56	SP80-0185	巴西 Brazil	115	VMC96-060	菲律宾 Philippines
57	SP80-1816	巴西 Brazil	116	VMC96-084	菲律宾 Philippines
58	SP80-3280	巴西 Brazil	117	VMC97-030	菲律宾 Philippines
59	SP81-3250	巴西 Brazil	118	VMC97-041	菲律宾 Philippines

(续表1) 试验种质名称及来源 Table 1(Continued) Name and origin of germplasm used in the experiment

1.2 基因组总DNA的提取

从每个材料取5个单株幼嫩叶片混合, 参照吴转娣等^{[ 15]}方法并略加改进提取基因组总DNA, 采用1%琼脂糖凝胶电泳检测提取DNA的质量和浓度, 将样品稀释到20 ng μL^-1, -20℃保存备用。

1.3 AFLP扩增及检测

根据Vos等^{[ 10]}方法进行适当调整, 采用本试验室开发的10对多态性较好的引物构建118份甘蔗种质的AFLP指纹图谱(表2)。基因组DNA经 EcoR I和 Mse I在37℃酶切14 h; 将酶切产物与 EcoR I和 Mse I特定接头在37℃连接2 h, 连接产物稀释10倍作为预扩增模板; EcoR I和 Mse I引物(含1个选择性核苷酸)预扩增的反应条件为94℃ 30 s, 56℃ 60 s, 72℃ 60 s, 20个循环。将预扩增产物稀释10倍作为选择性扩增模板, 以含3个选择性核苷酸的 EcoR I和 Mse I引物进行选择性扩增, PCR程序为94℃ 30 s, 65℃ 30 s, 72℃ 60 s, 1个循环, 然后以每循环复性温度逐级降低0.7℃的梯度继续12个循环, 变性和延伸条件同上, 最后以94℃ 30 s, 56℃ 30 s, 72℃ 60 s, 23个循环结束。所用PCR仪为德国Eppendorf Master Cycle Gradient。选择性扩增产物经95℃变性10 min后在5%的变性聚丙烯酰胺凝胶上电泳分离并参照刘新龙等^{[ 16]}的快速银染法染色。

表2 10对引物组合扩增的AFLP条带 Table 2 Amplification bands with ten pairs of AFLP primer

1.4 统计分析

AFLP扩增条带在相同迁移位置, 有带记为1, 无带记为0。标记位点的多态性条带比率(percentage of polymorphic bands, PPB), PPB= a/( a+ b), 标记位点的多态信息量(polymorphism information content, PIC)按公式PIC=1-∑ p_i 2计算; 标记系统的有效等位基因数(effective number of allele, Ne)按公式 Ne=1/∑ p_i 2计算, p_i表示 i位点的基因频率, 使用POPGEN32软件分析以上数据。同时使用NTSYS pc-V. 2.1计算相似系数, 即 Gs_ij= a/( a+ b+ c), Gs_ij是用来衡量2个体 i和 j的遗传相似性系数的, a为2个体的共享片段数, b和 c为 i个体和 j个体各自拥有的多态片段数。根据相似性系数进行UPGMA聚类分析(un-weighted pair group method analysis)和PCA主效应分析(principal component analysis), 并构建聚类图和主效应图。

2 结果与分析

2.1 AFLP扩增条带的多态性

采用10对本试验室开发的引物扩增118份参试材料, 扩增片段的大小主要在30~600 bp范围内, 且分布均匀, 共扩增出1310条带, 其中多态性条带1195条, 多态性比率为91.2%; 每个位点的平均多态信息量为0.2332, 每个位点的平均有效等位基因数为1.3789。从AFLP扩增条带的数量看, 中国材料(1189条) >菲律宾材料(1077条) >美国材料(1076条) >巴西材料(1003条) >法国材料(927条) >澳大利亚材料(888条); 从多态性比率看, 中国材料(86.8%) >美国材料(81%) >巴西材料(79.1%) >菲律宾材料(73.6%) >法国材料(69.9%) >澳大利亚材料(67.9%)。说明本研究筛选的10对引物多态性较高(表2)。

2.2 遗传相似性分析

参试的118份种质中, 遗传相似性系数在0.583~ 0.929之间, 平均为0.750。遗传相似性系数最高的是RB85-5113和SP80-0185, 达到0.929; 遗传相似性系数最低的是云蔗04-622和CP89-2376, 为0.583。从各国遗传相似性系数看, 澳大利亚材料(0.801)>菲律宾材料(0.775) >法国材料(0.768)>中国材料(0.753)>巴西材料(0.745)>美国材料(0.725)。说明各国种质之间的遗传基础差异较小, 亲缘关系较近, 其中美国种质遗传多样性相对较丰富。

2.3 聚类分析

根据遗传相似性系数, 用UPGMA法对118份甘蔗种质进行聚类分析(图1)。从图1可知, 在相似性系数0.69处切割时, 可划分为2个类群, 第I类群有3份种质材料, 包括粤糖00-236、云蔗04-622和SP80-1816, 这3份种质较早地与其他种质分开, 杂交组合选配中重点考虑; 第II类群有115份种质。在相似性系数0.74处切割时, 可将第II类群划分为5个亚群(A、B、C、D和E)。A亚群包含5份中国种质, 分别是ROC16、ROC22、云蔗94-375、云蔗89-7和云蔗94-343; B亚群包含12份美国和22份菲律宾种质; C亚群包含4份美国和19份澳大利亚种质; D亚群包含15份法国种质; E亚群包含26份中国和12份巴西种质。聚类结果表明, 大部分种质亲缘关系较近, 遗传组成相似; 美国种质在聚类中相对较分散, 说明在这118份种质中美国种质多样性相对较丰富; 第I类群和第II类群A亚群共计8份种质较早地和其他110份种质分开, 其中6份种质在中国已作为主要亲本在应用, 并得到了部分优良后代材料, 这6份种质分别是粤糖00-236、ROC16、ROC22、云蔗94-375、云蔗89-7和云蔗94-343; 其他2份种质云蔗04-622和SP80-1816在中国杂交育种中未见应用, 云蔗04-622突出特点为产量高和宿根性强, SP80- 1816为蔗糖分高和抗逆性强, 在杂交中应给予重点关注。

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	图1 118份甘蔗种质材料的AFLP聚类图Fig. 1 Dendrogram of 118 accessions of sugarcane germplasm with AFLP markers

2.4 主效应分析

基于Jaccard系数用PCA法对118份甘蔗种质AFLP标记结果进行主效应分析, 显示了不同种质的分类位置(图2)。22份菲律宾种质和12份美国种质位置集中在一起; 15份法国种质位置集中在一起, 这个区域还零星分布1份巴西和6份中国种质; 27份中国和12份巴西种质位置集中在一起; 19份澳大利亚种质和4份美国种质位置集中在一起。分子主效应分析结果与分子聚类结果一致, 集中在一个区域的种质亲缘关系较为紧密。

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	图2 甘蔗种质AFLP主效应分析图■:澳大利亚种质;▲:巴西种质;□:中国种质;●:法国种质;○:菲律宾种质; △: 美国种质。Fig. 2 Principal components analysis of sugarcane germplasm using AFLP data■: Australia germplasm; ▲: Brazil germplasm; □: China germplasm; ●: France germplasm; ○: Philippines germplasm; △: USA germplasm.

3 讨论

3.1 AFLP标记在甘蔗遗传多样性研究中的有效性

AFLP标记多态性强, 结果稳定, 重复性好, 是一种理想、有效的遗传标记。本研究采用AFLP标记技术对118份国内外甘蔗种质材料进行遗传多样性分析, 用10对自己筛选的引物扩增共得到1310条谱带, 其中多态性条带数目为1195条, 多态性条带比率为91.2%, 这低于庄南生等^{[ 8]}用4对AFLP引物对54份甘蔗种质扩增所得98.5%的多态性比率, 但远高于劳方业等^{[ 17]}用15对AFLP引物对78份中国自育甘蔗种质亲本扩增所得79.3%的多态性比率。多态性存在一定差异, 可能与引物数量、不同品种材料有一定关系, 但都显现出较高的多态性检出率。聚类结果则表明大部分种质亲缘关系较近, 遗传组成相似。通过对AFLP分子标记数据的相似性分析、主效应分析, 有效准确地反映出各国不同种质间的亲缘关系, 与目前种质材料之间普遍共祖现象相吻合。因此AFLP分子标记适合于各国甘蔗种质遗传多样性研究。

3.2 引进甘蔗种质的应用

甘蔗种质的引进是最简单、最经济、最有效的育种方法。一方面用于生产应用, 另一方面可用作亲本或中间材料。近年来, 引进的种质F134、ROC1、ROC10、ROC 16、ROC 22和ROC 25在我国甘蔗生产上取到了非常重要的作用, 尤其是ROC22取到了举足轻重的作用。甘蔗种质的引进和利用, 在中国甘蔗种质创新、甘蔗杂交育种和蔗糖产业发展中起到了重大作用。许多种质都曾经是我国的主栽品种, 目前ROC 22仍然占我国甘蔗栽培面积的60%左右。据不完全统计, 我国利用引进种质Co419、CP49-50、NCo310^{[ 1]}、CP72-1210^{[ 2]}, 分别育成了24、26、27和12个优良品种。桂糖11、ROC 10、ROC 16和ROC 22以及新良种福农94-0403、福农95-1702、桂糖25、桂糖26、粤糖93-159、粤糖00-236、云蔗94-375和云蔗99-596等均含有国外引进种质血缘。本研究从各国的遗传相似性系数看, 美国种质最小, 平均为0.725, 且美国种质在聚类中相对较分散, 表明美国种质遗传多样性相对较丰富, 在杂交利用中应给予关注。在聚类结果中, 我国主栽品种ROC 22分在II类群中的A亚群, 较早地和第II类群中的110份种质分开, 说明在遗传组成上和这110份种质有较大差异; 目前我国甘蔗杂交育种中每年用ROC 22为亲本配制的杂交组合多达200个以上, 且有优良后代材料如柳城03-182和柳城05-136等, 但ROC 22黑穗病较重, 目前的重点是如何利用、改良ROC 22, 为甘蔗产业作出更大的贡献。国外引进甘蔗种质在我国甘蔗产业中起到了重要作用, 在加强自主创新的同时, 应高效率应用好国外引进甘蔗种质, 尤其应加强将这些种质作为杂交亲本。

3.3 AFLP聚类与甘蔗种质系谱的关系

甘蔗系谱是杂交组合选配的重要依据。由于甘蔗杂交存在自交串粉等现象, 导致部分系谱不能真实反映其亲缘关系, 且甘蔗遗传背景复杂, 相同组合后代也可能存在较大遗传差异^{[ 8, 9]}。刘家勇等^{[ 9]}采用AFLP标记技术对68份甘蔗种质的亲缘关系研究显示, 来自同一杂交组合NCO310×QN54-7096的Q121、Q124、Q135和Q141, 只有Q121和Q124分在同一组, Q135和Q141被分在不同组中; 庄南生等^{[ 8]}采用AFLP标记技术对54份甘蔗种质研究表明, 来自同一杂交组合CP49-50×F134的广西1号和闽糖70-611, 并未分在一组。本研究中云蔗04-621、云蔗04-622和云蔗04-649都来自同一杂交组合云蔗89-7×崖城84-125, 其中只有云蔗04-621和云蔗04-649分在第II类群中的E亚群, 云蔗04-622分在第I类群, 这3个种质的母本云蔗89-7则分在第II类群中的A亚群。因此, 从分子水平揭示种质的遗传差异, 可为杂交组合选配中亲本的选择提供系谱数据之外的佐证依据。

4 结论

118份种质资源的遗传基础差异较小, 亲缘关系较近。各国遗传相似性系数中, 澳大利亚材料(0.801) >菲律宾材料(0.775) >法国材料(0.768) >中国材料(0.753) >巴西材料(0.745) >美国材料(0.725), 表明美国种质遗传多样性相对较丰富。聚类结果显示, 粤糖00-236、ROC16、ROC22、云蔗94-375、云蔗89-7、云蔗94-343、04-622和SP80-1816等8份种质较早地和其他110份种质分开, 其中粤糖00-236、ROC16、ROC22、云蔗94-375、云蔗89-7和云蔗94-343等6份种质已在中国作为主要亲本应用, 并得到了部分优良后代材料, 云蔗04-622和SP80- 1816在中国杂交育种中未见应用, 在杂交组合选配中应给予重点关注。

The authors have declared that no competing interests exist.

作者已声明无竞争性利益关系。The authors have declared that no competing interests exist.

参考文献

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[1]	李奇伟, 陈子云, 梁洪. 现代甘蔗改良技术. 广州: 华南理工大学出版社, 2000. pp11-18 Li Q W, Chen Z Y, Liang H. Modern Technology for Sugarcane Improvement. Guangzhou: South China University of Technology Press, 2000. pp11-18(in Chinese) [本文引用:2]
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[4]	Selvi A, Nair N V, Noyer J L, Singh N K, Balasundaram N, Bansal K C, Koundal K R, Mohapatra T. AFLP analysis of the phenetic organization and genetic diversity in the sugarcane complex, Saccharum and Erianthus. Genet Resour Crop Evol, 2006, 53: 831-842 [本文引用:2] [JCR: 1.593]
[5]	Harvey M, Huckett B I, Botha F C. Use of Polymerase Chain Reaction (PCR), and Rand om Amplification of Polymorphic DNAs, RAPDs, for the determination of genetic distances between 21 sugarcane varieties. In: Proceeding of South African Sugar Technologists Association, 1994, 68: 36-40 [本文引用:1]
[6]	Nair N V, Selvi A, Sreenivasan T V, Pushpalatha K N. Molecular diversity in Indian sugarcane cultivars as revealed by rand omly amplified DNA polymorphisms. Euphytica, 2002, 127: 219-225 [本文引用:1] [JCR: 1.643]
[7]	Lima M L A, Garcia A A F, Oliveira K M, Matsuoka S, Arizono H, Souza C L J, Souza A P. Analysis of genetic similarity detected by AFLP and coefficient of parentage among genotypes of sugar-cane (Saccharum spp. ). Theor Appl Genet, 2002, 104: 30-38 [本文引用:1] [JCR: 3.658]
[8]	庄南生, 郑成木, 黄东益, 唐燕琼, 高和琼. 甘蔗种质遗传基础的AFLP分析. 作物学报, 2005, 31: 444-450 Zhuang N S, Zheng C M, Huang D Y, Tang Y Q, Gao H Q. AFLP analysis for sugarcane germplasms. Acta Agron Sin, 2005, 31: 444-450 (in Chinese with English abstract) [本文引用:5] [CJCR: 1.667]
[9]	刘家勇, 赵培方, 刘新龙, 赵俊, 杨昆, 吴才文, 应雄美, 昝逢刚, 陈学宽. 68份国外甘蔗种质资源遗传多样性的AFLP分析. 湖南农业大学学报, 2013, 39: 466-470 Liu J Y, Zhao P F, Liu X L, Zhao J, Yang K, Wu C W, Ying X M, Zan F G, Chen X K. Genetic diversity analysis on 68 foreign sugarcane germplasms (Saccharum spp. ) with AFLP technique. J Hunan Agric Univ, 2013, 39: 466-470 (in Chinese with English abstract) [本文引用:3]
[10]	Vos P, Hogers R, Bleeker M, Reijans M, van de Lee T, Hornes M, Frijters A, Pot J, Peleman J, Kuiper M, Zabeau M. AFLP: a new technique for DNA fingerprinting. Nucl Acids Res, 1995, 23: 4407-4414 [本文引用:2]
[11]	劳方业, 刘睿, 何慧怡, 邓海华, 陈仲华, 陈健文, 符成, 张垂明, 杨业后. 崖城系列甘蔗亲本遗传多样性的AFLP标记分析. 分子植物育种, 2008, 6: 517-522 Lao F Y, Liu R, He H Y, Deng H H, Chen Z H, Chen J W, Fu C, Zhang C M, Yang Y H. AFLP analysis of genetic diversity in series sugarcane parents developed at HSBS. Mol Plant Breed, 2008, 6: 517-522 (in Chinese with English abstract) [本文引用:1] [CJCR: 1.23]
[12]	Schenck S, Crepeau M W, Wu K K, Moore P H, Yu Q, Ming R. Genetic diversity and relationships in native Hawaiian Saccharum officinarum sugarcane. J Hered, 2004, 95: 327-331 [本文引用:1] [JCR: 1.995]
[13]	李鸣, 谭裕模, 李杨瑞, 李容柏, 高国庆. 甘蔗(Saccharum officinarum L. )品种遗传差异的AFLP分子标记分析. 作物学报, 2004, 30: 1008-1013 Li M, Tan Y M, Li Y R, Li R B, Gao G Q. AFLP molecular analysis of genetic difference between cultivars in sugarcane (Saccharum officinarum L). Acta Agron Sin, 2004, 30: 1008-1013 (in Chinese with English abstract) [本文引用:1] [CJCR: 1.667]
[14]	刘新龙, 毛钧, 陆鑫, 马丽, Karen S A, Phillip A J, 蔡青, 范源洪. 甘蔗SSR和AFLP分子遗传连锁图谱构建. 作物学报, 2010, 36: 177-183 Liu X L, Mao J, Lu X, Ma L, Karen S A, Phillip A J, Cai Q, Fan Y H. Construction of molecular genetic linkage map of sugarcane based on SSR and AFLP markers. Acta Agron Sin, 2010, 36: 177-183 (in Chinese with English abstract) [本文引用:1] [CJCR: 1.667]
[15]	吴转娣, 昝逢刚, 赵丽宏, 罗遵喜, 张树珍. 甘蔗基因组DNA小量提取与大量提取方法研究. 生物技术通报, 2009, (增刊-1): 172-175 Wu Z D, Zan F G, Zhao L H, Luo Z X, Zhang S Z. Study on mini and mass-extraction methods of sugarcane genomic DNA. Biotechnol Bull, 2009, (suppl-1): 172-175 (in Chinese with English abstract) [本文引用:1] [CJCR: 0.49]
[16]	刘新龙, 蔡青, 毕燕, 陆鑫, 马丽, 应雄美, 毛钧. 甘蔗AFLP标记和SSR标记的PAGE胶快速银染检测方法. 江苏农业学报, 2009, 25: 433-435 Liu X L, Cai Q, Bi Y, Lu X, Ma L, Ying X M, Mao J. A rapid silver staining method for PAGE used in sugarcane AFLP and SSR molecular markers. Jiangsu J Agric Sci, 2009, 25: 433-435 (in Chinese with English abstract) [本文引用:1] [CJCR: 0.895]
[17]	劳方业, 刘睿, 何慧怡, 邓海华, 李奇伟, 陈仲华, 陈健文, 符成, 齐永文, 张垂明. 我国甘蔗亲本遗传多样性的AFLP标记分析. 基因组学与应用生物学, 2009, 28: 503-508 Lao F Y, Liu R, He H Y, Deng H H, Li Q W, Chen Z H, Chen J W, Fu C, Qi Y W, Zhang C M. Genetic diversity analysis of sugarcane parents with AFLP in China. Genom Appl Biol, 2009, 28: 503-508 (in Chinese with English abstract) [本文引用:1] [CJCR: 0.7323]

2000

0.0

... 目前, 甘蔗种质直接用于杂交利用较为普遍, 且作用巨大, 国外种质资源在我国杂交育种中也具有更重要的作用, 我国利用引进种质Co419、CP49-50、NCo310^[1]和CP72-1210^[2], 分别育成了24、26、27和12个优良品种 ...

... 据不完全统计, 我国利用引进种质Co419、CP49-50、NCo310^[1]、CP72-1210^[2], 分别育成了24、26、27和12个优良品种 ...

2007

0.0

... 据不完全统计, 我国利用引进种质Co419、CP49-50、NCo310^[1]、CP72-1210^[2], 分别育成了24、26、27和12个优良品种 ...

2006

0.0

... 表型性状极易受环境因素影响而变化, 仅依靠表型性状难以准确反映不同材料间的遗传差别和亲缘关系^[3,4] ...

... 由于AFLP标记多态性强, 结果稳定, 重复性好, 是一种理想、有效的遗传标记, 近年被广泛用于农作物的遗传多样性、遗传图谱、基因定位、指纹图谱等研究^[3,4,11,12] ...

2006

1.593

0.0

Selvi

, Nair

N V

, Noyer

J L

, Singh

N K

, Balasundaram

, Bansal

K C

, Koundal

K R

, Mohapatra

. AFLP analysis of the phenetic organization and genetic diversity in the sugarcane complex, Saccharum and Erianthus. Genet Resour Crop Evol, 2006, 53: 831-842

Amplified fragment length polymorphism (AFLP) markers were evaluated for determining the phylogenetic relationships, and the diversity in the Saccharum complex using 30 clones belonging to S. officinarum, S. robustum, S. spontaneum, S. barberi, S. sinense and the related genus Erianthus . The phenetic tree of the species clones based on AFLP data was consistent with the known taxonomical relationships. AFLP gave higher resolution of closely related species into discrete groups than that by RAPD and RFLP markers, reported earlier. The levels of diversity within the various Saccharum species were also found to be higher than those obtained previously with the same set of clones using RAPD markers. The intraspecies similarity in S. barberi and S. sinens e was much higher than interspecies similarity suggesting a clear separation of the two, which are considered ‘horticultural species’. The genetic similarity matrix derived from a single primer combination highly correlated ( r = 0.980) with that obtained from all the 12 primer combination used in the study, thus highlighting the efficiency of a single primer combination in delineating species relationships. All the primer combinations could identify markers that are specific to each of the species and the genus Erianthus . Among the species, specific markers were highest in S. spontaneum followed by S. robustum, S. barberi, S. officinarum and S. sinense . Erianthus had a distinct profile with 30% of the total amplified fragments being specific to it. This offers great scope for identifying intergeneric hybrids, which has been very difficult using morphological traits and RAPD markers. High degree of correspondence between the results from the cluster analysis based on Jaccard's similarity index, Neighbour Joining tree based on Sokal and Michener distance matrix and AFTD (Analyses Factorielle on Table of Distances) analysis clearly demonstrated that AFLP markers would be an appropriate tool in providing better information about the relationships among the species, estimation of diversity, and in revealing species and genus specific markers that could be directly applied in sugarcane breeding programmes.

1.Indian Agricultural Research Institute National Research Centre on Plant Biotechnology New Delhi 110012 India New Delhi 110012 India 2.Sugarcane Breeding Institute Coimbatore 641007 Tamilnadu India Coimbatore 641007 Tamilnadu India 3.CIRAD UMR 1096 PIA, TA 40/03 F-34398 Montpellier France UMR 1096 PIA, TA 40/03 F-34398 Montpellier France

... 表型性状极易受环境因素影响而变化, 仅依靠表型性状难以准确反映不同材料间的遗传差别和亲缘关系^[3,4] ...

1994

0.0

... 品种系谱虽然在一定程度上能反映品种之间的亲缘关系, 但由于自交、串粉等因素, 存在一定的局限性^[5,6,7], 基于分子水平的亲缘关系较基于系谱的亲缘关系具有更高的可靠性^[8,9] ...

2002

1.643

0.0

Nair

N V

, Selvi

, Sreenivasan

T V

, Pushpalatha

K N

. Molecular diversity in Indian sugarcane cultivars as revealed by rand omly amplified DNA polymorphisms. Euphytica, 2002, 127: 219-225

Genetic diversity in 28 prominent Indian sugarcane varieties cultivated under a wide range of agroclimatic conditions, was studied using 25 RAPD markers. The mean genetic distance among the 28 varieties was only 29.31%, implying that a large part of the genome is similar among the varieties. This probably arises from the lack of parental diversity, with few clones which are themselves related, contributing to the parentage of these varieties. The parentage of the varieties did not contribute significantly to the clustering pattern. Varieties belonging to the same parentage were grouped under different clusters while varieties from different parentages were grouped under the same cluster. The tropical and subtropical identities of the varieties also did not contribute to the clustering pattern as individual clusters included varieties from both tropics and subtropics. This shows that genetically similar varieties are present in both the regions. Among the varieties, Co 7717 was found to be totally distinct and divergent from rest of the varieties. The study reveals the limited genetic base of the current Indian commercial varieties and the need to diversify the genetic base by using new sources from the germplasm.

1.Sugarcane Breeding Institute, Coimbatore Crop Improvement Division 641 007 Tamil Nadu India 641 007 Tamil Nadu India

2002

3.658

0.0

Lima M L A, Garcia A A F, Oliveira

K M

, Matsuoka

, Arizono

, Souza C L J, Souza

A P

. Analysis of genetic similarity detected by AFLP and coefficient of parentage among genotypes of sugar-cane (Saccharum spp. ). Theor Appl Genet, 2002, 104: 30-38

Despite the economical importance of sugar cane, until the present-date no studies have been carried out to determine the correlation of the molecular-based genetic similarity (GS) and the coefficient of parentage ( f )-estimates generated for cultivars. A comprehensive knowledge of the amount of genetic diversity in parental cultivars, could improve the effectiveness of breeding programmes. In this study, amplified fragment length polymorphism (AFLP) and pedigree data were used to investigate the genetic relationship in a group of 79 cultivars (interspecific hybrids), used as parents in one of the Brazilian breeding programmes, and four species of Saccharum ( Saccharum sinense , Saccharum barberi and two of Saccharum officinarum ) . The objectives of this study were to assess the level of genetic similarity among the sugar-cane cultivars and to investigate the correlation between the AFLP-based GS and f, based on pedigree information. Twenty one primer combinations were used to obtain the AFLP molecular markers, generating a total of 2,331 bands, of which 1,121 were polymorphic, with a polymorphism rate, on average, of 50% per primer combination. GSs were determined using Jaccard’s similarity coefficient, and a final dendrogram was constructed using an unweighted pair-group method using arithmetic average (UPGMA). AFLP-based GS ranged from 0.28 to 0.89, with a mean of 0.47, whereas f ranged from 0 to 0.503, with a mean of 0.057. Cluster analysis using GS divided the genotypes into related subgroups suggesting that there is important genetic relationship among the cultivars. AFLP-based GS and f were significantly correlated ( r = 0.42, P r value suggests that the AFLP data may help to more-accurately quantify the degree of relationship among sugar-cane cultivars.

1.Centro de Biologia Molecular e Engenharia Genética (CBMEG), Universidade Estadual de Campinas (UNICAMP), Cidade Universitária ”Zeferino Vaz”, CP 6010, CEP 13083-970, Campinas, SP, Brasil e-mail: anete@unicamp.br Tel.: (55-19) 3788-1132 or 3788-1130, Fax: (55-19) 3788-1089, Present address: A.P. de Souza, Centro de Biologia Molecular e Engenharia Genética (CBMEG), Universidade Estadual de Campinas (UNICAMP), Cidade Universitária ”Zeferino Vaz”, CP 6010, CEP 13083-970, Campinas, SP, Brasil BR 2.Departamento Genética e Evolu??o – Instituto de Biologia – Universidade Estadual de Campinas (UNICAMP), Cidade Universitária ”Zeferino Vaz”, CP 6109, CEP 13083-970, Campinas, SP, Brasil BR 3.Departamento de Ciências Exatas, Escola Superior de Agricultura ”Luiz de Queiroz” (ESALQ), Universidade de S?o Paulo (USP), CP 9, CEP 13400-970, Piracicaba, SP, Brasil BR 4.Departamento de Biotecnologia Vegetal (DBV), Centro de Ciências Agrárias (CCA), Universidade Federal de S?o Carlos (UFSCar), CEP 13600-970, Araras, SP, Brasil BR 5.Departamento de Genética, Escola Superior de Agricultura ”Luiz de Queiroz” (ESALQ), Universidade de S?o Paulo (USP), CP 83, CEP 13400-970, Piracicaba, SP, Brasil BR

2005

0.0

1.667

庄南生, 郑成木, 黄东益, 唐燕琼, 高和琼. 甘蔗种质遗传基础的AFLP分析. 作物学报, 2005, 31: 444-450

Zhuang

N S

, Zheng

C M

, Huang

D Y

, Tang

Y Q

, Gao

H Q

. AFLP analysis for sugarcane germplasms. Acta Agron Sin, 2005, 31: 444-450 (in Chinese with English abstract)

Modern sugarcane cultivars (Saccharum spp.) are polyploid or aneuploid clones with complex genetic background, and the major components of the genome derived from S. officinarum, and the remainder from S. spontaneum, S. barberi, S. robustum, S. sinense, and Erianthus arundinaceus. In this study, 54

College of Agronomy, South China University of Tropical Agriculture, Danzhou 571737, Hainan

采用AFLP分子标记技术对54份甘蔗种质（14份祖亲种、40份栽培品种或品系）的遗传基础进行了分析。利用筛选出的4对多态性较强的引物组合（M+CAG/E+ACG，M+CTC/E+ACT，M+CTG/E+ACC，M+CTG/E+ACG），构建了甘蔗54份种质的AFLP指纹图谱，这4对引物组合共扩增出396条谱带，其中多态带390条，占98.5%。54份种质的遗传相似系数变化

... 庄南生等^[8]采用AFLP分子标记技术分析54份甘蔗种质(14份祖亲种、40份栽培品种或品系)的遗传基础, 利用筛选出的4对多态性较强的引物组合, 构建了甘蔗54份种质的AFLP指纹图谱 ...

... 2%, 这低于庄南生等^[8]用4对AFLP引物对54份甘蔗种质扩增所得98 ...

... 由于甘蔗杂交存在自交串粉等现象, 导致部分系谱不能真实反映其亲缘关系, 且甘蔗遗传背景复杂, 相同组合后代也可能存在较大遗传差异^[8,9] ...

... 庄南生等^[8]采用AFLP标记技术对54份甘蔗种质研究表明, 来自同一杂交组合CP49-50#cod#x000d7 ...

2013

0.0

... 由于甘蔗杂交存在自交串粉等现象, 导致部分系谱不能真实反映其亲缘关系, 且甘蔗遗传背景复杂, 相同组合后代也可能存在较大遗传差异^[8,9] ...

... 刘家勇等^[9]采用AFLP标记技术对68份甘蔗种质的亲缘关系研究显示, 来自同一杂交组合NCO310#cod#x000d7 ...

1995

0.0

... AFLP标记技术综合了RFLP和RAPD的优点, 既具有RFLP的稳定性和PCR反应快速、灵敏的特点, 又具有RAPD的方便性, 同时还克服了RFLP和RAPD的缺点^[10] ...

... 3 AFLP扩增及检测根据Vos等^[10]方法进行适当调整, 采用本试验室开发的10对多态性较好的引物构建118份甘蔗种质的AFLP指纹图谱(表2) ...

2008

0.0

1.23

劳方业, 刘睿, 何慧怡, 邓海华, 陈仲华, 陈健文, 符成, 张垂明, 杨业后. 崖城系列甘蔗亲本遗传多样性的AFLP标记分析. 分子植物育种, 2008, 6: 517-522

Lao

F Y

, Liu

, He

H Y

, Deng

H H

, Chen

Z H

, Chen

J W

, Fu

, Zhang

C M

, Yang

Y H

. AFLP analysis of genetic diversity in series sugarcane parents developed at HSBS. Mol Plant Breed, 2008, 6: 517-522 (in Chinese with English abstract)

采用15对多态性较好的AFLP引物对52份崖城系列甘蔗亲本品系的遗传多样性进行分析,结果表明,每对引物的多态性位点平均为61.40,多态性位点率为77.30%.52份材料的遗传相似系数在0.483 2～0.961 4之间,平均为0.680 6,遗传多样性属中等水平.来自同一组合"CP72-1210×崖城82-108"品系的遗传相似性系数很高,在0.783 6～0.839 8范围内,平均是0.811 7.以遗传相似系数阈值约为0.670 0划分,聚类分析把52份材料分为5大类,系谱记录中亲缘关系密切的品系,大多数都能归为同一类.不同年代品系的遗传多样性指数差异明显,最高是20世纪90年代,为0.315 5,最低是20世纪的50～70年代,为0.265 4;来自相一组合的后代品系遗传多样性指数较低.建议在亲本创新过程中,同一亲本不宜过多重复使用.

2004

1.995

0.0

2004

0.0

1.667

李鸣, 谭裕模, 李杨瑞, 李容柏, 高国庆. 甘蔗(Saccharum officinarum L. )品种遗传差异的AFLP分子标记分析. 作物学报, 2004, 30: 1008-1013

, Tan

Y M

, Li

Y R

, Li

R B

, Gao

G Q

. AFLP molecular analysis of genetic difference between cultivars in sugarcane (Saccharum officinarum L). Acta Agron Sin, 2004, 30: 1008-1013 (in Chinese with English abstract)

5 pairs of AFLP primers from 64 pairs were selected, which had rich polymorphism. The genetic diversity of two sugarcane varieties was analyzed with the fingerprinting mapped by the 5 primer pairs，and the genetic similarity and distance were estimated. The results showed that there were distinct di

Guangxi Crop Genetic Improvement and Biotechnology Laboratory，Nanning 530007, Guangxi

采用AFLP技术，从64对引物中筛选出5对带型分布均匀、多态性丰富且分辨能力强的引物，对两个在生产上广泛推广应用的甘蔗品种的AFLP指纹图谱进行分析，计算了两品种间的遗传相似性和遗传距离。结果表明，5对引物在2个甘蔗品种间均存在显著的差异，其中多态性位点占总扩增位点的10.2%，区分率达100%。这对应用AFLP技术鉴定甘蔗

... 李鸣等^[13]采用AFLP技术, 用5对引物扩增在生产上广泛推广的2个甘蔗品种, AFLP指纹图谱分析区分率达100% ...

2010

0.0

1.667

刘新龙, 毛钧, 陆鑫, 马丽, Karen

S A

, Phillip

A J

, 蔡青, 范源洪. 甘蔗SSR和AFLP分子遗传连锁图谱构建. 作物学报, 2010, 36: 177-183

Liu

X L

, Mao

, Lu

, Ma

, Karen

S A

, Phillip

A J

, Cai

, Fan

Y H

. Construction of molecular genetic linkage map of sugarcane based on SSR and AFLP markers. Acta Agron Sin, 2010, 36: 177-183 (in Chinese with English abstract)

To take advantage of the MAS technique and develop the molecular genetic linkage map in sugarcane breeding program in China, we constructed two molecular genetic maps by single dose markers (SDM) and double dose markers (DDM) from 36 SSR and 12 AFLP primers for two mapping populations (F 1 and BC 1 ). F 1 population was from cross combination of sugarcane commercial variety (Co419) × S. spontaneum (Y75/1/2) consisting of 269 true individuals, BC 1 population from backcross combination of F 1 02/356×sugarcane commercial variety (ROC25) with 266 true individuals. All plants from two populations were detected by PCR. Two molecular genetic linkage maps were established. 298 SDMs and DDMs were acquired by c 2 test from 630 segregation markers of F 1 population, about 47% of total markers; 264 SDMs and DDMs were obtained from 571 segregation markers of BC 1 population which were 46% of total markers, Co419>02/356>Y75/1/2>ROC25 in efficiency of getting SDMs and DDMs. Based on LOD ≥ 5.0, genetic distance ≤ 40 cM between markers, 134 of 298 SDMs and DDMs from F 1 population formed 55 linkage groups (LGs), among which 39 LGs were included in eight homology groups(HGs), 16 LGs were excluded, the map length reached 1 458.3 cM with an average distance of 10.9 cM between markers; 133 of 298 SDMs and DDMs from BC 1 population made up 47 LGs , among which 34 LGs were composed in eight HGs except for 13 LGs, the map covered 1 059.6 cM with an average distance of 8.0 cM. At the same time, these SDMs and DDMs from Co419 and Y75/1/2 formed 34 LGs and 20 LGs, respectively, and these SDMs from two parents only made up 1 LGs; 19 LGs were obtained from these SDMs and DDMs of 02/356, and ROC25, 9 LGS were formed by SDMs from two parents; Co419>Y75/1/2>02/356=ROC2 according to forming number of LGs. These results indicated that F 1 mapping population was better than BC 1 mapping population in quality of constructing linkage map. The maps will provide the valuable information for locating gene, further research on genetic linkage map and MAS breeding.

1 Yunnan Key Laboratory of Sugarcane Genetic Improvement, Kaiyuan 661600, China; 2 Sugarcane Research Institute, Yunnan Academy of Agricultural Sciences, Kaiyuan 661600, China; 3 Biotechnology & Genetic Resources Institute, Yunnan Academy of Agricultural Sciences, Kunming 650223, China; 4 CSIRO Division of Plant Industry, Brisbane, QLD 4067, Australia

采用甘蔗商业品种 Co419 与野生种割手密 Y75/1/2 杂交，获得 269 个单株，组成 F 1 群体，用 F 1 02/356 与商业品种 ROC25 回交获得 266 个单株，组成 BC 1 群体。利用筛选的多态性条带丰富的 36 对 SSR 引物和 12 对 AFLP 引物，对两个群体进行 PCR 扩增和分子遗传连锁分析，构建甘蔗分子遗传连锁图谱。用 F 1 群体获得 630 个分离标记，经 c 2 检测， 298 个标记为单双剂量标记，占总标记数的 47% ；用 BC 1 群体获得 571 个分离标记，有 264 个标记为单双剂量标记，占总标记数的 46% ； 4 个亲本获得单双剂量标记的数量依次为 Co419>02/356>Y75/1/2>ROC25 。在 LOD ≥ 5.0 ，相邻标记遗传距离≤ 40 cM 的条件下， F 1 群体有 134 个单双剂量标记被纳入 55 个连锁群，其中 39 个连锁群归属 8 个同源组， 16 个未列入，总遗传距离为 1 458.3 cM ，标记间平均图距为 10.9 cM ； BC 1 群体有 133 个单双剂量标记被纳入 47 个连锁群，其中 34 个连锁群归属于 8 个同源组， 13 个连锁群未列入，总遗传距离为 1 059.6 cM ，标记间平均图距为 8.0 cM 。从 4 个亲本单双剂量标记进入的连锁群数来看， Co419 最多，归入 34 个连锁群，其次为 Y75/1/2 ，归入 20 个连锁群，第 3 为 02/356 和 ROC25 ，归入 19 个连锁群。研究结果表明 , 从单双剂量标记比例、形成连锁群数量、总遗传距离来看， F 1 群体构图质量要优于 BC 1 群体。

... 刘新龙等^[14]采用SSR和AFLP标记技术, 对构建的2个群体进行PCR扩增和分子遗传连锁分析,构建了甘蔗分子遗传连锁图谱 ...

2009

0.0

0.49

吴转娣, 昝逢刚, 赵丽宏, 罗遵喜, 张树珍. 甘蔗基因组DNA小量提取与大量提取方法研究. 生物技术通报, 2009, (增刊-1): 172-175

Z D

, Zan

F G

, Zhao

L H

, Luo

Z X

, Zhang

S Z

. Study on mini and mass-extraction methods of sugarcane genomic DNA. Biotechnol Bull, 2009, (suppl-1): 172-175 (in Chinese with English abstract)

甘蔗的叶片中含有大量的多糖、多酚类和RNA等物质,对甘蔗分子生物学实验带来了极大的影响.本研究利用改良的CTAB对甘蔗基因组DNA进行小量提取法与大量提取方法的研究,本方法操作简单,DNA完整性好、产量高、纯度高可满足大部分分子实验的需要.

... 2 基因组总DNA的提取从每个材料取5个单株幼嫩叶片混合, 参照吴转娣等^[15]方法并略加改进提取基因组总DNA, 采用1%琼脂糖凝胶电泳检测提取DNA的质量和浓度, 将样品稀释到20 ng #cod#x003bc ...

2009

0.0

0.895

刘新龙, 蔡青, 毕燕, 陆鑫, 马丽, 应雄美, 毛钧. 甘蔗AFLP标记和SSR标记的PAGE胶快速银染检测方法. 江苏农业学报, 2009, 25: 433-435

Liu

X L

, Cai

, Bi

, Lu

, Ma

, Ying

X M

, Mao

. A rapid silver staining method for PAGE used in sugarcane AFLP and SSR molecular markers. Jiangsu J Agric Sci, 2009, 25: 433-435 (in Chinese with English abstract)

银染方法是一种重要的DNA染色法.其原理是利用银离子和核苷酸结合,在碱性环境下甲醛使银离子还原从而使凝胶中的DNA片段得以显现[1,2].

... 选择性扩增产物经95℃变性10 min后在5%的变性聚丙烯酰胺凝胶上电泳分离并参照刘新龙等^[16]的快速银染法染色 ...

2009

0.0

0.7323

劳方业, 刘睿, 何慧怡, 邓海华, 李奇伟, 陈仲华, 陈健文, 符成, 齐永文, 张垂明. 我国甘蔗亲本遗传多样性的AFLP标记分析. 基因组学与应用生物学, 2009, 28: 503-508

Lao

F Y

, Liu

, He

H Y

, Deng

H H

, Li

Q W

, Chen

Z H

, Chen

J W

, Fu

, Qi

Y W

, Zhang

C M

. Genetic diversity analysis of sugarcane parents with AFLP in China. Genom Appl Biol, 2009, 28: 503-508 (in Chinese with English abstract)

采用15对多态性较好的AFLP引物对我国自育的78个甘蔗亲本材料遗传多样性进行分析,结果表明,每对引物的多态性位点平均为65.67,多态性位点率为8255%.78个甘蔗亲本的遗传相似系数在0.4535～0.8927之间,平均为0.6821 .相同组合后代的遗传相似系数较高,平均为0.765 6.以遗传相似系数阈值约为0.700划分,聚类分析把78个亲本分为7大类,系谱记录中亲缘关系密切的亲本品系,大多数都能归为同一类.遗传多样性指数分析显示,不同年代亲本多样性指数差异明显,20世纪80年代最高,为0.318 5,70年代最低,为0.264 5;不同省区的亲本遗传多样指数变化更显著,在0.195 2～0.299 9之间,广东最高,云南最低.

... 5%的多态性比率, 但远高于劳方业等^[17]用15对AFLP引物对78份中国自育甘蔗种质亲本扩增所得79 ...