白冬梅^,1,*, 薛云云¹, 赵姣姣², 黄莉², 田跃霞¹, 权宝全¹, 姜慧芳^,2,*1山西省农业科学院经济作物研究所, 山西汾阳 032200
2中国农业科学院油料作物研究所, 湖北武汉 430062

Identification of Cold-tolerance During Germination Stage and Genetic Diversity of SSR Markers in Peanut Landraces of Shanxi Province

BAI Dong-Mei^,1,*, XUE Yun-Yun¹, ZHAO Jiao-Jiao², HUANG Li², TIAN Yue-Xia¹, QUAN Bao-Quan¹, JIANG Hui-Fang^,2,* 1 Industrial Crops Research Institute, Shanxi Academy of Agricultural Sciences, Fenyang 032200, Shanxi, China
2 Oil Crops Research Institute, Chinese Academy of Agricultural Sciences, Wuhan 430062, Hubei, China

通讯作者: ^* 通信作者(Corresponding authors): 白冬梅, E-mail: baidm1221@163.com; 姜慧芳, E-mail: peanut@oilcrops.com, Tel: 027-86711550

收稿日期:2018-02-24接受日期:2018-06-12网络出版日期:2018-07-03

基金资助:

本研究由国家现代农业产业技术体系建设专项.CARS-13 山西省农业科学院生物育种工程项目.17yzgc051 山西省农业科学院科技自主创新能力提升工程项目.2017zzlx-07

Received:2018-02-24Accepted:2018-06-12Online:2018-07-03

Fund supported:

This study was supported by the China Agricultural Research System.CARS-13 the Biological Breeding Project of Shanxi Academy of Agricultural Sciences.17yzgc051 the Science and Technology Independent Innovation Ability Enhancement Project of Shanxi Academy of Agricultural Sciences.2017zzlx-07

摘要
低温寒害是引起花生产量和品质下降的主要因素之一, 培育和种植高产稳产耐寒性强的品种是降低低温寒害的理想途径。然而高耐寒性种质的缺乏和耐寒性鉴定的困难, 是限制耐寒性育种取得突破的主要原因。本研究对72份山西花生地方品种进行芽期耐寒性鉴定, 以其相对发芽率和相对发芽指数作为耐寒性评价指标, 初步将其分为高耐寒、耐寒、中感、敏感、高感5级。为了解山西花生地方品种耐寒性遗传多样性, 合理、高效利用耐寒型花生资源, 用多态性好的90对SSR引物评价了不同耐寒性花生品种, 结果显示, 参试品种遗传多样性存在较大差异, 在遗传距离为0.4时, 被聚类为三大类群。3份高耐寒品种和7份耐寒品种分别聚类到不同的3个类群中, 说明耐寒性花生遗传多样性丰富。
关键词： 花生;地方品种;芽期耐寒性;遗传多样性

Abstract
Cold injury is one of the main factors causing yield and quality decline in peanut. Cultivating and planting varieties with high and stable yield and strong cold tolerance is an ideal way to reduce cold injury, However, the lack of high cold tolerant germplasm and the difficulty of cold tolerance identification are the main reasons to limit the breakthrough of cold tolerance breeding. In this study, 72 local peanut varieties in Shanxi province were identified for cold tolerance at germination stage. Based on their relative germination rate and relative germination index, the cold tolerance of 72 peanut cultivars we preliminarily divided the 72 peanut cultivars into five grades, namely high-cold-tolerant type, cold-tolerant type, middle type, sensitive type, and high sensitive type. Ninety pairs of SSR primers with good polymorphism were used to evaluate the peanut cultivars with different cold-tolerance levels, and to examine the genetic diversity of cold-tolerant peanut landraces in Shanxi province making rational and efficient use of cold tolerant peanut resources. The tested cultivars were highly different in genetic diversity and were clustered into three groups with genetic distance of 0.4. Three high-cold-tolerant cultivars and seven cold-tolerant cultivars were clustered into three different groups, indicating that the cold-tolerant peanut varieties are rich in genetic diversity.
Keywords：peanut;landraces;cold-tolerance at germination stage;genetic diversity


PDF (1933KB)元数据多维度评价相关文章导出EndNote|Ris|Bibtex收藏本文
本文引用格式
白冬梅, 薛云云, 赵姣姣, 黄莉, 田跃霞, 权宝全, 姜慧芳. 山西花生地方品种芽期耐寒性鉴定及SSR遗传多样性[J]. 作物学报, 2018, 44(10): 1459-1467. doi:10.3724/SP.J.1006.2018.01459
BAI Dong-Mei, XUE Yun-Yun, ZHAO Jiao-Jiao, HUANG Li, TIAN Yue-Xia, QUAN Bao-Quan, JIANG Hui-Fang. Identification of Cold-tolerance During Germination Stage and Genetic Diversity of SSR Markers in Peanut Landraces of Shanxi Province[J]. Acta Agronomica Sinica, 2018, 44(10): 1459-1467. doi:10.3724/SP.J.1006.2018.01459


花生(Arachis hypogaea L.)是我国重要的油料和经济作物, 在国内大宗油料作物中, 单位面积产量、产油量、种植效益以及国际市场竞争力等均具有明显优势^[1]。目前国内食用油过度依赖进口, 油脂供给安全问题凸显, 进一步发展我国花生生产, 拓宽花生种植区域, 是满足不断增长的市场需求、提高农业生产效益、增加农民收入的迫切需要^[2]。然而, 在花生播种后常常遭遇低温寒害, 种子活力受到损害, 出苗率明显降低, 轻者延缓花生萌发和幼苗生长发育, 重者发生大面积低温烂种, 缺苗断垄, 导致严重减产。高耐寒性种质的缺乏和耐寒性鉴定的困难, 是限制耐寒性育种取得突破的主要原因之一。

山西省地处黄土高原, 大部分地区海拔在1500 m以上, 年平均气温3~14℃。山西省花生种植历史悠久, 经过漫长的自然训化和人工选择, 孕育了丰富的变异类型和较强的抗逆性^[3]。我们对山西省地方品种的农艺性状和品质性状详细分析表明, 山西省花生地方品种具有丰富的遗传多样性^[4,5]。本研究对72份山西省地方花生品种进行了芽期耐寒性鉴定及SSR多态性检测, 分析研究其耐寒性遗传多样性, 为花生耐寒性育种及其相关研究提供了丰富的遗传资源和重要的理论依据。

1 材料与方法

1.1 参试花生品种

本试验选取山西省农业科学院经济作物研究所花生课题组征集的72份山西花生地方品种, 其中普通型51份、多粒型11份、珍珠豆型10份, 其编号、品名、类型见表1。

Table 1
表1
表172份山西供试花生地方品种编号、品名、类型
Table 1Landrace type and origin of 72 peanuts from Shanxi province

编号 Code	品种 Cultiver	植物学类型 Botanical type	编号 Code	品种 Cultiver	植物学类型 Botanical type
1	汾西小粒Fenxixiaoli	珍珠豆型vulgaris	37	洪洞花生4 Hongtonghuasheng 4	普通型hypogaea
2	襄汾油花生Xiangfenyouhuasheng	多粒型fastigiata	38	洪洞大粒Hongtongdali	普通型hypogaea
3	隰县一把抓Xixianyibazhua	普通型hypogaea	39	乡宁花生Xiangninghuasheng	普通型hypogaea
4	洪洞花生5 Hongtonghuasheng 5	多粒型fastigiata	40	临汾小粒Linfenxiaoli	普通型hypogaea
5	难山小粒 Nanshanxiaoli	珍珠豆型vulgaris	41	临汾一窝蜂Linfenyiwofeng	多粒型fastigiata
6	吉县大花生Jixiandahuasheng	普通型hypogaea	42	高平花生Gaopinghuasheng	普通型hypogaea
7	大宁大花生Daningdahuasheng	普通型hypogaea	43	黎城花生Lichenghuasheng	普通型hypogaea
8	太谷二粒Taiguerli	珍珠豆型vulgaris	44	长子花生Zhangzihuasheng	多粒型fastigiata
9	石楼小粒Shilouxiaoli	珍珠豆型vulgaris	45	运城花生 Yunchenghuasheng	普通型hypogaea
10	临县多粒Linxianduoli	珍珠豆型vulgaris	46	新绛大花生Xinjiangdahuasheng	普通型hypogaea
11	安泽落花生Anzeluohuasheng	多粒型fastigiata	47	永济爬地垄Yongjipadilong	普通型hypogaea
12	吉县大粒秧Jixiandaliyang	普通型hypogaea	48	平陆大粒Pingludali	普通型hypogaea
13	翼城花生Yichenghuasheng	珍珠豆型vulgaris	49	稷山花生Jishanhuasheng	普通型hypogaea
14	浮山一把抓Fushanyibazhua	普通型hypogaea	50	榆次花生Yucihuasheng	普通型hypogaea
15	中阳花生Zhongyanghuasheng	普通型hypogaea	51	榆次伏花生Yucifuhuasheng	普通型hypogaea
16	沁水花生Qinshuihuasheng	普通型hypogaea	52	灵石小花生Lingshixiaohuasheng	珍珠豆型vulgaris
17	黎城花生Lichenghuasheng	普通型hypogaea	53	太谷大粒Taigudali	普通型hypogaea
18	运城小角花Yunchengxiaojiaohuasheng	普通型hypogaea	54	阳曲花生Yangquhuasheng	普通型hypogaea
19	运城大蔓花生Yunchengdamanhuasheng	普通型hypogaea	55	文水大花生Wenshuidahuasheng	普通型hypogaea
20	稷山小蔓Jishanxiaoman	普通型hypogaea	56	文水多粒Wenshuiduoli	多粒型fastigiata
21	垣曲长蔓Yuanquchangman	普通型hypogaea	57	汾阳大粒Fenyangdali	普通型hypogaea
22	祁县小花生Qixianxiaohuasheng	珍珠豆型vulgaris	58	汾阳多粒Fenyangduoli	多粒型fastigiata
23	兴县大花生Xingxiandahuasheng	普通型hypogaea	59	汾阳四粒红Fenyangsilihong	多粒型fastigiata
24	文水花生Wenshuihuasheng	普通型hypogaea	60	柳林二粒Liulinerli	普通型hypogaea
25	孝义花生Xiaoyihuasheng	普通型hypogaea	61	榆社花生Yushehuasheng	珍珠豆型vulgaris
26	临县大粒Linxiandali	普通型hypogaea	62	榆社红花生Yushehonghuasheng	多粒型fastigiata
27	临县小粒Linxianxiaoli	普通型hypogaea	63	榆社大粒Yushedali	普通型hypogaea
28	交口花生Jiaokouhuasheng	普通型hypogaea	64	曲沃一窝蜂Quwoyiwofeng	多粒型fastigiata
29	汾阳小粒Fenyangxiaoli	普通型hypogaea	65	洪洞花生1 Hongtonghuasheng 1	普通型hypogaea
30	柳林花生Liulinhuasheng	普通型hypogaea	66	武乡花生Wuxianghuasheng	普通型hypogaea
31	侯马大粒 Houmadali	普通型hypogaea	67	武乡白花生Wuxiangbaihuasheng	珍珠豆型vulgaris
32	吉县大粒1 Jixiandali 1	普通型hypogaea	68	武乡黑花生Wuxiangheihuasheng	普通型hypogaea
33	吉县大粒蔓Jixiandaliman	普通型hypogaea	69	武乡多粒Wuxiangduoli	多粒型fastigiata
34	曲沃小花生Quwoxiaohuasheng	普通型hypogaea	70	武乡彩粒Wuxiangcaili	普通型hypogaea
35	曲沃一把抓Quwoyibazhua	普通型hypogaea	71	河曲大粒Hequdali	普通型hypogaea
36	大宁一把抓Daningyibazhua	普通型hypogaea	72	柳林小粒Liulinxiaoli	普通型hypogaea

新窗口打开|下载CSV

1.2 芽期耐寒性鉴定

采用BIC-300人工气候箱模拟大田气象条件, 进行芽期耐寒性筛选鉴定。选取2016年和2017年成熟饱满、种皮完整、大小一致的种子20粒, 采用纸间发芽, 3次重复, 常温浸种8 h后放入12℃人工气候箱培养72 h, 然后2℃低温胁迫暗培养96 h, 再调到10℃暗培养72 h, 最后置于25℃人工气候箱恒温发芽, 以常温浸种8 h后在25℃发芽为对照, 每天计录发芽种子数, 18 d后计算各品种的相对发芽率(%)和相对发芽指数(GI), GI = ∑(Gt/Dt)。Gt为第t天的发芽种子数, Dt为相对应的发芽日数。

1.3 SSR引物及PCR扩增

本研究在中国农业科学院油料作物研究所花生生物技术实验室进行, 所用的154对SSR引物由该实验室提供。选取花生健壮幼叶, 采用优化的CTAB法^[6]提取基因组DNA, 用1.0%琼脂糖凝胶电泳检测DNA质量, 紫外分光光度计测定其浓度并统一调整DNA浓度至100 ng L^-1。PCR反应体系为10 μL, 含Mix 2.5 μL (由北京全式金生物技术有限公司生产)、ddH₂O 5 μL、10~40 pmol L^-1引物对0.5 μL、10~20 ng模板DNA 2 μL。扩增条件为94℃预变性3 min; 93℃变性30 s, 55~65℃ (不同引物退火温度不同)退火30 s, 72℃延伸1 min, 共32个循环; 72℃延伸10 min。PCR产物经6%变性聚丙烯酰胺凝胶电泳检测, 硝酸银染色, 显影, 扫描保存。

1.4 数据统计分析

根据PCR扩增结果, 以0、1、C统计SSR扩增带型, 在相同迁移率位置上, 有带记为“1”, 无带记为“0”, 缺失记为“C”, 建立相应的数据库, 用Microsoft Excel 2013处理基本数据。再根据不同分析软件的格式要求作相应转换。用Powermarker-V 3.25 软件^{[7,8,9,10,11]}计算每对引物的多样性参数, 包括等位基因数(Na)、主基因频率(MAF)、基因多样性指数(H)、多态性信息含量指数(PIC), 利用非加权组平均法(UPGMA)进行聚类分析, 生成聚类图; 用Popgene Ver.1.32 分析Shannon’s信息指数(I)。

2 结果与分析

2.1 芽期耐寒性品种分析

吸胀后突遇低温胁迫, 各种质材料间表现出明显的差异, 有的材料受低温影响小, 能正常发芽, 生长状况良好。有的材料耐低温能力很差, 低温胁迫后, 种子霉烂, 不能正常发芽生长。其他材料介于这两种类型之间, 萌发情况参差不齐(图1)。

图1

新窗口打开|下载原图ZIP|生成PPT
图1低温胁迫后部分花生品种萌发情况

Fig. 1Germination of part peanut seeds under low temperature stress



由表2可见, 72份材料相对发芽率变幅范围为6.86%~96.50%, 相对发芽指数变幅范围为12.87%~ 94.46%, 表现最好的临县多粒、新降大花生、榆次花生3份材料的相对发芽率与相对发芽指数均>90%, 可作为高耐寒材料; 而表现最差的2份材料侯马大粒和文水多粒的相对发芽率低于10%, 相对发芽指数低于20%, 可作为高感材料。按照相对发芽率和相对发芽指数的变幅范围, 将参试的72份材料耐寒性分为5级。一级为高耐寒材料, 相对发芽率与相对发芽指数均>90%, 有3份, 占总材料的4.17%, 其中珍珠豆型1份, 普通型2份; 二级为耐寒材料, 相对发芽率>90%, 80%<相对发芽指数<90%, 有7份, 占总材料的9.72%, 其中珍珠豆型1份, 多粒型3份, 普通型3份; 三级为中间材料, 50%<相对发芽率<90%, 50%<相对发芽指数<80%, 有17份, 占总材料的23.61%, 其中有珍珠豆型1份, 多粒型2份, 普通型14份; 四级为敏感材料, 10%<相对发芽率<50%, 20%<相对发芽指数<50%, 有43份, 占总材料的59.72%, 其中珍珠豆型7份, 多粒型5份, 普通型31份; 五级为高感材料, 相对发芽率<10%, 相对发芽指数<20%, 有2份, 占总材料的2.78%, 其中多粒型1份, 普通型1份。四级和五级材料受低温胁迫, 种子霉烂, 活力丧失, 不能正常发芽生长, 严重影响出苗, 形成田间缺苗断垄影响产量。鉴定筛选出的高耐寒和耐寒材料中, 包括2份珍珠豆型, 3份多粒型, 5份普通型, 说明花生耐寒性与品种植物学属性关系不大。

Table 2
表2
表2供试品种的耐寒性鉴定
Table 2Identification of cold-tolerant the cultivars in the present study

基因型 Gene type	编号 Code	相对发芽率 Relative germination rate (%)			相对发芽指数 Relative germination index (%)
		2016	2017	Mean	2016	2017	Mean
高耐寒	10	92.27	94.44	93.36	93.01	91.29	92.15
High cold-tolerant	46	96.64	96.36	96.50	94.20	94.71	94.46
	50	94.64	93.10	93.87	90.15	94.03	92.09
耐寒	1	94.44	92.72	93.58	86.14	85.09	85.62
Cold-tolerant	2	90.60	92.30	91.45	81.47	81.70	81.59
	24	94.64	92.47	93.56	85.93	86.23	86.08
	44	98.56	92.72	95.64	81.42	86.81	84.12
	47	94.34	92.98	93.66	87.55	88.87	88.21
	63	93.57	92.72	93.15	87.07	89.59	88.33
	64	98.18	94.23	96.21	84.00	82.08	83.04
中间	9	64.78	76.47	70.63	51.18	53.20	52.19
Middle	15	55.12	60.34	57.73	51.69	51.45	51.57
	23	71.69	68.52	70.11	55.64	54.34	54.99
	25	51.77	54.54	53.16	53.91	55.52	54.72
	32	86.50	78.42	82.46	62.95	61.72	62.34
	33	86.60	78.97	82.79	68.64	62.15	65.40
	34	77.35	76.36	76.86	55.80	55.15	55.48
	36	62.71	74.54	68.63	52.54	52.14	52.34
	37	70.33	66.67	68.50	50.73	52.31	51.52
	38	77.35	70.59	73.97	52.35	54.28	53.32
	39	63.00	55.36	59.18	50.13	50.20	50.17
	40	88.47	79.25	83.86	51.72	57.93	54.83
	41	59.22	64.81	62.02	50.27	53.71	51.99
	49	82.95	80.36	81.66	58.19	58.06	58.13
	57	54.70	50.00	52.35	51.67	52.61	52.14
	60	55.12	52.63	53.88	50.33	50.68	50.51
	69	73.01	72.22	72.62	51.90	60.75	56.33
基因型 Gene type	编号 Code	相对发芽率 Relative germination rate (%)			相对发芽指数 Relative germination index (%)
		2016	2017	Mean	2016	2017	Mean
敏感	3	28.55	31.47	30.01	33.36	31.33	32.35
Sensitive	4	31.41	32.73	32.07	40.62	34.00	37.31
	5	48.11	37.50	42.81	42.97	40.00	41.49
	6	43.18	43.63	43.41	31.59	29.32	30.46
	7	16.67	21.81	19.24	36.85	27.72	32.29
	8	29.27	29.82	29.55	31.91	32.17	32.04
	11	41.18	33.33	37.26	20.61	23.27	21.94
	12	40.37	38.89	39.63	36.89	34.07	35.48
	13	30.68	29.63	30.16	23.95	24.78	24.37
	14	33.93	41.81	37.87	49.76	47.43	48.60
	16	36.01	33.93	34.97	38.36	35.65	37.01
	17	28.84	27.78	28.31	35.59	34.87	35.23
	18	30.80	32.73	31.77	23.17	27.10	25.14
	19	29.41	32.68	31.05	35.57	33.08	34.33
	20	22.88	28.84	25.86	37.53	38.47	38.00
	21	22.22	21.82	22.02	29.78	28.45	29.12
	22	26.39	25.92	26.16	25.62	27.28	26.45
	26	37.71	35.71	36.71	23.47	23.72	23.60
	27	36.19	36.84	36.52	31.32	29.26	30.29
	28	35.69	29.82	32.76	20.67	21.30	20.99
	29	35.90	34.55	35.23	21.16	21.12	21.14
	30	49.95	46.55	48.25	27.91	30.77	29.34
	35	41.18	31.48	36.33	34.88	31.58	33.23
	42	39.34	33.93	36.64	31.05	32.78	31.92
	43	47.51	48.28	47.90	40.62	39.76	40.19
	45	21.05	38.18	29.62	21.64	25.45	23.55
	48	39.64	33.33	36.49	29.29	30.76	30.03
	51	29.67	30.35	30.01	28.36	25.78	27.07
	52	26.32	21.43	23.88	25.14	24.19	24.67
	53	16.67	30.90	23.79	36.80	34.28	35.54
	54	28.79	29.82	29.31	19.77	19.86	19.82
	55	24.97	36.36	30.67	20.66	22.69	21.68
	58	48.23	49.13	48.68	32.72	38.17	35.45
	59	24.53	29.82	27.18	40.78	34.90	37.84
	61	35.69	34.55	35.12	35.37	34.58	34.98
	62	15.79	15.25	15.52	36.80	21.39	29.10
	65	33.37	35.71	34.54	29.35	30.18	29.77
	66	32.64	30.90	31.77	29.48	23.86	26.67
	67	43.18	47.26	45.22	34.22	28.71	31.47
	68	33.33	33.93	33.63	27.82	31.45	29.64
	70	36.19	33.90	35.05	25.21	26.44	25.83
	71	48.23	50.00	49.12	37.68	37.61	37.65
	72	16.08	21.82	18.95	30.50	30.58	30.54
高感	31	8.20	11.76	9.98	18.20	16.90	17.55
High-sensitive	56	6.82	6.90	6.86	16.09	9.64	12.87

新窗口打开|下载CSV

2.2 SSR标记多态性分析

选用8份不同地理来源、不同类型的花生品种基因组DNA, 用154对SSR引物进行PCR扩增。有101对引物扩增出稳定、清晰可辨的谱带, 有效扩增比率为47.3%, 其中90对具有多态性, 多态性引物比率为89.1% (图2)。最终确定了谱带清晰稳定且具多态性的90对SSR引物用于本研究。

图2

新窗口打开|下载原图ZIP|生成PPT
图2引物A06B209在72份花生品种中的扩增

Fig. 2Amplified bands using primer A06B209 in 72 peanuts materials



从表3可以看出, 72份山西省地方品种在90个SSR标记中扩增出317个等位基因数(Na), 每对引物平均扩增出3.5222个等位基因, 不同引物每个位点等位基因数差异很大, 变化范围2~8个, 检测到的等位基因最多的引物是A07B516和A08A90A。主基因频率(MAF)变化幅度为0.2917 (A07B34)~ 0.8986 (A02B62), 平均0.6834。基因多样性指数(GD)变化幅度为0.1823 (A02B62)~0.7711 (A08A90), 平均0.4537; 多态信息含量指数(PIC)变异范围为0.1657 (A02B62)~0.7378 (A08A90), 平均0.4047; Shannon’s信息指数(I)变异范围为0.3283 (A02B62)~ 1.6734 (A08A90), 平均0.8092。表明本研究所用的SSR位点的遗传多样性丰富, 显示山西省花生地方品种遗传多样性方面存在较大差异。其中16对引物最有效, Shannon’s信息指数均在1以上, 综合比较各参数, 等位基因数值多的SSR位点, 基因多样性指数、多态信息含量指数和Shannon’s信息指数的值也大, 三者变化趋势一致, 因此, 基因多样性指数、多态信息含量指数和Shannon’s信息指数对于遗传多样性分析, 更具有可靠的实际意义。

Table 3
表3
表390个SSR标记的遗传参数
Table 3Genetic parameters of 90 SSR markers used in this study

项目 Item	等位基因数Allele number (Na)	主基因频率 Major allele frequency (MAF)	基因多样性指数 Gene diversity (GD)	多态信息含量指数 Polymorphism information content (PIC)	Shannon’s信息指数 Shannon’s information index (I)
最大值 Max	8	0.8986	0.7711	0.7378	1.6734
最小值 Min	2	0.2917	0.1823	0.1657	0.3283
合计 Total	317
平均值 Mean	3.5222	0.6834	0.4537	0.4047	0.8092

新窗口打开|下载CSV

2.3 不同耐寒性花生品种的遗传多样性

根据SSR标记数据, 利用Powermarker-V3.25软件, 采用非加权组平均法(UPGMA)聚类分析表明, 72份参试材料在遗传距离为0.4时, 被分为三大类群。第I类群是以多粒型为主的10个品种, 其中包括2份耐寒材料; 第II类是以普通型为主51个品种, 其中包括2份高耐寒材料和3份耐寒材料; 第III类是以珍珠豆型为主的11个品种, 其中包括1份高耐寒材料和2份耐寒材料, 说明花生耐寒性遗传多样性丰富(图3)。耐寒品种汾西小粒和长子花生亲缘关系最远, 而高耐寒品种新降大花生和高感品种侯马大粒亲缘关系最近, 说明高耐品种和高感品种并不是亲缘关系最远, 耐寒品种间也不是亲缘关系最近。

图3

新窗口打开|下载原图ZIP|生成PPT
图3基于SSR标记的72份山西花生地方品种的聚类分析图

Fig. 3Dendrogram of 72 peanut landraces in Shanxi province based on SSR markers

3 讨论

花生芽期寒害是引起花生产量和品质下降的主要因素之一, 山西地处黄土高原高海拨区, 其地方品种具有丰富的遗传多样性, 可能蕴含着高耐寒型基因。因此, 本研究以山西花生地方品种为试验材料, 采用人工气候箱中模拟大田气象条件, 可以不受季节气候条件等因素的限制而开展鉴定工作, 从而加快鉴定进度。而且人工气候箱设备的培养条件(光照、温度、水分等)易于控制且重复性好, 鉴定出的耐寒性花生品种在田间更具有适应性。本研究中耐寒性鉴定设置了2℃低温胁迫, 这是经过多次的、几个温度梯度的试验得出来的结果, 温度低于2℃, 多数试材会被淘汰, 温度高会出现大部分材料的正常发芽出苗而达不到选择的目的, 这与封海胜^[12]所做的花生种子吸胀期间耐低温性鉴定是一致的。刘海龙等^[13]利用花生种质资源耐低温表型方法鉴定花生种质资源耐低温属性。吕建伟等^[14]以花生相对出苗率将花生种质资源划分为不耐、低耐、中耐、高耐4个等级。唐月异等^[15]以露白率及芽长/种长作为鉴定花生吸胀期耐寒性的指标。目前对于花生不同阶段耐寒性评价没有统一的标准, 本研究结合前人研究基础, 把相对发芽率和相对发芽指数作为鉴定花生品种芽期耐寒性的指标, 初步将72份花生品种耐寒性分为高耐寒、耐寒、中感、敏感、高感5级。本研究鉴定筛选出的高耐寒和耐寒材料中, 包括2份珍珠豆型、3份多粒型和5份普通型, 表明花生耐寒性与品种植物学属性关系不大, 这与唐月异等^[15]研究得出的结论一致。常硕其等^[16]对杂交品种亲本及后代进行耐寒性鉴定时, 发现水稻的耐寒性是可以通过杂交稳定遗传的, 后代与亲本的耐寒性呈正相关。康旭梅等^[17]提出, 在综合考虑后代优良性状的同时, 只要保证亲本之一的耐寒性较强, 就有可能保证F₁耐寒性强且具有优良的综合性状。这些研究足以证明筛选耐寒性资源作为杂交亲本培育耐寒性杂交后代的育种方法是完全可行的。本研究筛选出的临县多粒、新降大花生、榆次花生等耐寒品种可以作为亲本进行杂交培养耐寒性后代, 为选育高产高油酸耐寒性强新品种及其相关研究提供材料基础和理论依据。

分子标记是揭示花生遗传多样性的有效手段, 国内外研究者曾用RAPD^[18]、SSR^{[19,20,21,22,23,24,25,26]}、AFLP^[27,28]等分子标记对花生种质资源遗传多样性广泛研究。其中, SSR 以共显性好、多态性丰富成为花生最实用的检测标记。本研究所用的SSR位点的遗传多样性丰富, 显示出山西省花生地方种质资源遗传多样性方面存在较大差异。综合比较各遗传参数, 等位基因数值多的SSR位点, 基因多样性指数(GD)、多态信息含量指数(PIC)和Shannon’s信息指数(I)的值也大, 三者变化趋势一致。因此, 基因多样性指数、多态信息含量指数和Shannon’s信息指数对于遗传多样性分析, 更具有可靠的实际意义, 这一结果与花生^[11]、豌豆^[10,29-30]和小扁豆^[31]上的研究结果一致。在遗传距离为0.4时, 参试品种被聚为三个类群, 类群I以多粒型为主, 类群II以普通型为主, 类群III是珍珠豆型为主, 说明SSR 标记的聚类与花生植物属性关系密切, 与花生耐寒性特性和地域特性关系不大, 3份高耐寒品种和7份耐寒品种分布于3个不同的类群中, 这可能与控制耐寒性的遗传因子在不同品种间存在差异有关。本研究所用材料仅限于山西省花生地方品种, 分析花生耐寒性遗传特性的分子基础, 还需要更多的验证分析和丰富的育种材料, 以取得更加可靠的结果, 从而为花生耐寒性育种及其相关研究提供理论依据和材料基础。

4 结论

通过对72份山西地方品种芽期耐寒性鉴定, 初步筛选出3份高耐寒品种和7份耐寒品种, 分布在三个不同的类群中, 说明花生耐寒品种遗传多样性丰富。

The authors have declared that no competing interests exist.

作者已声明无竞争性利益关系。

参考文献 View Option 原文顺序
文献年度倒序
文中引用次数倒序
被引期刊影响因子

[1] 禹山林 . 中国花生品种及其系谱. 上海: 上海科学技术出版社, 2008. pp 1-5
[本文引用: 1]

Yu S L. Peanut Varieties and Pedigree in China. Shanghai: Shanghai Scientific and Technical Publishers, 2008. pp 1-5(in Chinese)
[本文引用: 1]

[2] 万书波 . 中国花生栽培学. 上海: 上海科学技术出版社, 20013. pp 20-21
[本文引用: 1]

Wan S B. Peanut Cultivation in China. Shanghai: Shanghai Scientific and Technical Publishers, 2013. pp 20-21(in Chinese)
[本文引用: 1]

[3] 沈浩, 刘登义 . 遗传多样性概述
生物学杂志, 2001,18(2):4-7
[本文引用: 1]

Shen H, Liu D Y . Summary of genetic diversity
Chin J Biol, 2001,18(2):4-7 (in Chinese with English abstract)
[本文引用: 1]

[4] 白冬梅, 王国桐, 薛云云, 田跃霞, 权宝全 . 山西省地方花生品种农艺性状的遗传多样性分析
山西农业科学, 2014,42:542-547
[本文引用: 1]

Bai D M, Wang G T, Xue Y Y, Tian Y X, Quan B Q . Analysis of genetic diversity of Shanxi peanut landrace based on agronomic traits
Shanxi Agric Sci, 2014,42:542-547 (in Chinese with English abstract)
[本文引用: 1]

[5] 白冬梅, 王国桐, 薛云云, 田跃霞, 权宝全 . 山西省地方花生种质资源品质性状的综合评价
中国农学通报, 2014,30(24):187-193
URL [本文引用: 1]
In order to explore the use value of 90 samples of peanut collected from Shanxi province and select high-quality varieties, this studymeasured and analyzed their quality traits in laboratory, including protein content, oil content, content of oleic acid and linoleic acid, and discussed the relationship between the quality traits and geographic origin and different groups. The results showed that the diversity indexes of five quality traits were 2.04, 2.02, 2.02, 1.99, 1.93, indicating that the quality traits of peanut germplasm resources in Shanxi showed rich genetic diversity. The coefficient variation of the 5 quality traits were 0.06, 0.09, 0.11, 0.11, 0.23, indicating that there was a significant difference between the quality traits among the resources. The 90 samples were divided into 4 categories by cluster analysis. High-quality resources were screened, for example Wuxiang red peanuts, Linxian multigerm, Yongji small wasp waist. The results provided a scientific theoretical basis for rational utilization of local germplasm resources and breeding of new peanut varieties with high quality in Shanxi province.
Bai D M, Wang G T, Xue Y Y, Tian Y X, Quan B Q . Comprehensive evaluation of peanut germplasm resources in Shanxi province based on quality traits
Chin Agric Sci Bull, 2014,30(24):187-193 (in Chinese with English abstract)
URL [本文引用: 1]
In order to explore the use value of 90 samples of peanut collected from Shanxi province and select high-quality varieties, this studymeasured and analyzed their quality traits in laboratory, including protein content, oil content, content of oleic acid and linoleic acid, and discussed the relationship between the quality traits and geographic origin and different groups. The results showed that the diversity indexes of five quality traits were 2.04, 2.02, 2.02, 1.99, 1.93, indicating that the quality traits of peanut germplasm resources in Shanxi showed rich genetic diversity. The coefficient variation of the 5 quality traits were 0.06, 0.09, 0.11, 0.11, 0.23, indicating that there was a significant difference between the quality traits among the resources. The 90 samples were divided into 4 categories by cluster analysis. High-quality resources were screened, for example Wuxiang red peanuts, Linxian multigerm, Yongji small wasp waist. The results provided a scientific theoretical basis for rational utilization of local germplasm resources and breeding of new peanut varieties with high quality in Shanxi province.

[6] Cuc L M, Mace E S, Crouch J H, Quang V D, Long T D, Varshney R K . Isolation and characterization of novel microsatellite markers and their application for diversity assessment in cultivated ground-nut (Arachis hypogaea L.)
BMC Plant Biol, 2008,8:55
[本文引用: 1]

[7] 陈本银, 姜慧芳, 廖伯寿, 任小平, 黄家权, 雷永, 王圣玉 . 野生花生种质的 SSR 遗传多样性
热带亚热带植物学报, 2008,16:296-303
DOI:10.3969/j.issn.1005-3395.2008.04.002URL [本文引用: 1]
以花生属(Arachis)6个区组24种(包括栽培种)84份种质为材料,用SSR技术对其亲缘关系和遗传多样性进行了分析。从206对SSR引物中筛选到59对能扩增出稳定的多态性条带的引物,这些引物能在花生属基因组DNA中扩增出1～6个DNA片段。结果表明,84份种质的遗传距离为0.04～0.93,平均为0.64,其中匍匐区组的A.appressipila的2份种质(G4与G5)的遗传距离最小(0.04),匍匐区组的A.rigonii(G14)与根茎区组的A.glabrata(G28)的遗传距离最大(0.93)。聚类分析结果与花生属的区组分类基本一致,栽培种花生被聚在花生区组中,而且7份栽培种被聚在同一亚亚组中,相同植物学类型(相当于变种)的材料均被分别聚在一起。异形花区组与直立区组的亲缘关系最近,与花生区组的亲缘关系较近的是匍匐区组。花生区组的二倍体野生种A.villosa、A.duranensis和A.benensis与栽培种花生关系较近,可以作为桥梁物种来转移其他野生花生的优良基因。
Chen B Y, Jiang H F, Liao B S, Ren X P, Huang J Q, Lei Y, Wang S Y . Genetic diversity analysis of Arachis gerplasm by SSR
J Trop Subtrop Bot, 2008,16:296-303 (in Chinese with English abstract)
DOI:10.3969/j.issn.1005-3395.2008.04.002URL [本文引用: 1]
以花生属(Arachis)6个区组24种(包括栽培种)84份种质为材料,用SSR技术对其亲缘关系和遗传多样性进行了分析。从206对SSR引物中筛选到59对能扩增出稳定的多态性条带的引物,这些引物能在花生属基因组DNA中扩增出1～6个DNA片段。结果表明,84份种质的遗传距离为0.04～0.93,平均为0.64,其中匍匐区组的A.appressipila的2份种质(G4与G5)的遗传距离最小(0.04),匍匐区组的A.rigonii(G14)与根茎区组的A.glabrata(G28)的遗传距离最大(0.93)。聚类分析结果与花生属的区组分类基本一致,栽培种花生被聚在花生区组中,而且7份栽培种被聚在同一亚亚组中,相同植物学类型(相当于变种)的材料均被分别聚在一起。异形花区组与直立区组的亲缘关系最近,与花生区组的亲缘关系较近的是匍匐区组。花生区组的二倍体野生种A.villosa、A.duranensis和A.benensis与栽培种花生关系较近,可以作为桥梁物种来转移其他野生花生的优良基因。

[8] Lin K . Muse S V. PowerMarker: an integrated analysis environment for genetic marker analysis
Bioinformatics, 2005,21:2128-2129
DOI:10.1093/bioinformatics/bti282URLPMID:15705655 [本文引用: 1]
PowerMarker delivers a data-driven, integrated analysis environment (IAE) for genetic data. The IAE integrates data management, analysis and visualization in a user-friendly graphical user interface. It accelerates the analysis lifecycle and enables users to maintain data integrity throughout the process. An ever-growing list of more than 50 different statistical analyses for genetic markers has been implemented in PowerMarker.

[9] Ferguson M E, Burow M D, Schulze S R, Bramel P J, Paterson A H, Kresovich S, Mitchell S . Microsatellite identification and characterization in peanut (A. hypogaea L.)
Theor Appl Genet, 2004,108:1064-1070
DOI:10.1007/s00122-003-1535-2URLPMID:15067392 [本文引用: 1]
A major constraint to the application of biotechnology to the improvement of the allotetraploid peanut, or groundnut ( Arachis hypogaea L.), has been the paucity of polymorphism among germplasm lines using biochemical (seed proteins, isozymes) and DNA markers (RFLPs and RAPDs). Six sequence-tagged microsatellite (STMS) markers were previously available that revealed polymorphism in cultivated peanut. Here, we identify and characterize 110 STMS markers that reveal genetic variation in a diverse array of 24 peanut landraces. The simple-sequence repeats (SSRs) were identified with a probe of two 27,648-clone genomic libraries: one constructed using Pst I and the other using Sau 3AI/ Bam HI. The most frequent, repeat motifs identified were ATT and GA, which represented 29% and 28%, respectively, of all SSRs identified. These were followed by AT, CTT, and GT. Of the amplifiable primers, 81% of ATT and 70.8% of GA repeats were polymorphic in the cultivated peanut test array. The repeat motif AT showed the maximum number of alleles per locus (5.7). Motifs ATT, GT, and GA had a mean number of alleles per locus of 4.8, 3.8, and 3.6, respectively. The high mean number of alleles per polymorphic locus, combined with their relative frequency in the genome and amenability to probing, make ATT and GA the most useful and appropriate motifs to target to generate further SSR markers for peanut.

[10] He G H, Meng R H, Newman M, Gao G Q, Pittman R N, Prakash C S . Microsatellites as DNA markers in cultivated peanut (Arachis hypogaea L.)
BMC Plant Biol, 2003,3:3
URL [本文引用: 2]

[11] 任小平, 张晓杰, 廖伯寿, 雷永, 黄家权, 陈玉宁, 姜慧芳 . ICRISAT 花生微核心种质资源SSR 标记遗传多样性分析
中国农业科学, 2010,43:2848-2858
[本文引用: 2]

Ren X P, Zhang X J, Liao B S, Lei Y, Huang J Q, Chen Y N, Jiang H F . Analysis of genetic diversity in ICRISAT mini core collection of peanut( Arachis hypogaea L.) by SSR markers
.Sci Agric Sin, 2010, 43:2848-2858 (in Chinese with English abstract)
[本文引用: 2]

[12] 封海胜 . 花生种子吸胀期间耐低温性鉴定
中国油料, 1991, ( 1):67-70
[本文引用: 1]

Feng H S . Identification on low temperature resistance of peanut seed during imbibition stage
Chin Oil Crop Sci, 1991, ( 1):67-70 (in Chinese)
[本文引用: 1]

[13] 刘海龙, 陈小姝, 杨富军, 白冬梅, 孙晓苹, 吕永超, 任小平, 姜慧芳, 高华援 . 花生种质资源耐低温表型鉴定方法研究
花生学报, 2017,46:20-25
DOI:10.14001/j.issn.1002-4093.2017.03.004URL [本文引用: 1]
本研究目的是构建既可鉴定花生种质资源耐低温属性,又可进行耐低温性数量性状分析的表型鉴定方法。本方法构建公式相对发芽率=xi/yi×100%,花生耐低温性属性分级标准鉴定花生耐低温属性,及利用相对发芽率数据进行耐低温数量性状分析。利用此方法对94份花生种质资源进行鉴定,鉴定出相对发芽率≥85%耐低温型（R）6个,约占检测资源的6.38%,分别为山花8（A1）、豫花1（A38）、开农30（A10）、白沙1016（A37）、油4（A32）和泉花646（A42）,从94份花生资源中选取20份材料进行田间耐低温验证,其结果与本研究构建的方法结果一致,证明花生种质资源耐低温表型方法可以鉴定花生种质资源耐低温属性;利用此方法鉴定出徐花13×中花6重组自交系群体（RIL）各家系及亲本,分析耐低温性主基因＋多基因遗传模型,符合模型G-1,主基因的遗传率为91.21%,多基因的遗传率为8.34%,证明此方法可应用在耐低温性数量性状分析上。
Liu H L, Chen X S, Yang F J, Bai D M, Sun X P, Lyu Y C, Ren X P, Jiang H F, Gao H Y . Research of identification method on low temperature resistance of peanut germpasm resources phenotype
J Peanut Sci, 2017,46:20-25 (in Chinese with English abstract)
DOI:10.14001/j.issn.1002-4093.2017.03.004URL [本文引用: 1]
本研究目的是构建既可鉴定花生种质资源耐低温属性,又可进行耐低温性数量性状分析的表型鉴定方法。本方法构建公式相对发芽率=xi/yi×100%,花生耐低温性属性分级标准鉴定花生耐低温属性,及利用相对发芽率数据进行耐低温数量性状分析。利用此方法对94份花生种质资源进行鉴定,鉴定出相对发芽率≥85%耐低温型（R）6个,约占检测资源的6.38%,分别为山花8（A1）、豫花1（A38）、开农30（A10）、白沙1016（A37）、油4（A32）和泉花646（A42）,从94份花生资源中选取20份材料进行田间耐低温验证,其结果与本研究构建的方法结果一致,证明花生种质资源耐低温表型方法可以鉴定花生种质资源耐低温属性;利用此方法鉴定出徐花13×中花6重组自交系群体（RIL）各家系及亲本,分析耐低温性主基因＋多基因遗传模型,符合模型G-1,主基因的遗传率为91.21%,多基因的遗传率为8.34%,证明此方法可应用在耐低温性数量性状分析上。

[14] 吕建伟, 马天进, 李正强, 陈锋, 姜慧芳 . 花生种质资源出苗期耐低温性鉴定方法及应用
花生学报, 2014,43(3):13-18
[本文引用: 1]

Lyu J W, Ma T J, Li Z Q, Chen F, Jiang H F . Identification method of low temperature tolerance on peanut germplasm resource
J Peanut Sci, 2014,43(3):13-18 (in Chinese with English abstract)
[本文引用: 1]

[15] 唐月异, 王传堂, 高华媛, 凤桐, 张树伟, 王秀贞, 张建成, 禹山林 . 花生种子吸胀期间耐低温性及其与品质性状的相关研究
核农学报, 2011,25:436-442
URL [本文引用: 2]
Tolerance to low temperaterature during germination for fifty-five accessions of peanut germplasm were evaluated when the seeds were imbibed at 2℃ at 96h followed by 25℃ for 72h. Percentage of the number of seeds with radicals breaking through testa(PSWRBT) and ratio of the length of hypocotyls and radicals to the length of seeds(RHRS) were calculated. Main quality traits including oleic acid, linoleic acid, palmitic acid, oil, protein and sucrose contents were determined by near-infrared spectroscopy. Relationship between low temperature tolerance of peanut seed during imbibition and individual quality attributes was analyzed. The results showed that under low temperature stress condition, 3,3,5 and 43 accessions had the PSERTBT of ≥80%, 70%~80%, 60%~70% and0.5, 0.4~0.5, 0.4~0.3 and
Tang Y Y, Wang C T, Gao H Y, Feng T, Zhang S W, Wang X Z, Zhang J C, Yu S L . Low temperature tolerance during seed imbibition and its relationship to main quality in peant
Acta Agric Nucl Sin, 2011, 25:436-442 (in Chinese with English abstract)
URL [本文引用: 2]
Tolerance to low temperaterature during germination for fifty-five accessions of peanut germplasm were evaluated when the seeds were imbibed at 2℃ at 96h followed by 25℃ for 72h. Percentage of the number of seeds with radicals breaking through testa(PSWRBT) and ratio of the length of hypocotyls and radicals to the length of seeds(RHRS) were calculated. Main quality traits including oleic acid, linoleic acid, palmitic acid, oil, protein and sucrose contents were determined by near-infrared spectroscopy. Relationship between low temperature tolerance of peanut seed during imbibition and individual quality attributes was analyzed. The results showed that under low temperature stress condition, 3,3,5 and 43 accessions had the PSERTBT of ≥80%, 70%~80%, 60%~70% and0.5, 0.4~0.5, 0.4~0.3 and

[16] 常硕其, 邓启云, 罗祎, 陈小龙 . 超级杂交稻及其亲本的耐冷性研究
杂交水稻, 2015,30(1):51-57
[本文引用: 1]

Chang S Q, Deng Q Y, Luo W, Chen X L . Research on low temperature tolerance in super hybrid rice and parent
Hybrid Rice, 2015,30(1):51-57 (in Chinese with English abstract)
[本文引用: 1]

[17] 康旭梅, 李小湘, 潘孝武, 刘文强 . 普通野生稻耐冷性研究进展
杂交水稻, 2017,32(2):1-5
DOI:10.16267/j.cnki.1005-3956.201702001URL [本文引用: 1]
普通野生稻是栽培稻的近缘种,含有许多特异基因,耐冷性突出,有利于耐冷性水稻品种的培育.综述了普通野生稻耐冷基因的定位与克隆、耐冷生理生化和分子机制及其在水稻耐冷性改良中的应用等研究进展,对今后普通野生稻耐冷性相关研究作了展望.
Kang X M, Li X X, Pan X W, Liu W Q . Research progress on low temperature tolerance in Oryza rufipogon Griff
Hybrid Rice, 2017, 32(2):1-5 (in Chinese with English abstract)
DOI:10.16267/j.cnki.1005-3956.201702001URL [本文引用: 1]
普通野生稻是栽培稻的近缘种,含有许多特异基因,耐冷性突出,有利于耐冷性水稻品种的培育.综述了普通野生稻耐冷基因的定位与克隆、耐冷生理生化和分子机制及其在水稻耐冷性改良中的应用等研究进展,对今后普通野生稻耐冷性相关研究作了展望.

[18] Subramanian V, Gurtu S, Rao R C N, Nigam S N . Identification of DNA polymorphism in cultivated groundnut using random amplified polymorphic DNA (RAPD) assay
Genome, 2000,43:656-660
DOI:10.1139/g00-034URL [本文引用: 1]

[19] Tang R H, Gao G Q, He L Q, Han Z Q, Shan S H, Zhong R C, Zhou C Q, Jiang J, Li Y R, Zhuang W J . Genetic diversity in cultivated groundnut based on SSR markers
J Genet Genomics, 2007,34:449-459
DOI:10.1016/S1673-8527(07)60049-6URLPMID:17560531 [本文引用: 1]
Peanut(Arachis hypogaea L.) is an important source crop for edible oil and protein. It is important to identify the genetic diversity of peanut genetic resources for cultivar development and evaluation of peanut accessions. Thirty-four SSR markers were used to assess the genetic variation of four sets of twenty-four accessions each from the four botanical varieties of the cultivated peanut. Among the tested accessions,ten to sixteen pairs of SSR primers showed polymorphisms. The maximum differentiation index,which was defined as the degree of genetic differentiation,was as high as 0.992 in the tested accessions. Each accession could be discriminated by a specific set of polymorphic SSR primers,and the intra-variety genetic distance was determined among accessions,with an average of 0.59 in var. fastigiata,0.46 in var. hypogaea,0.38 in var. vulgaris,and 0.17 in var. hirsuta. Dendro-grames based on genetic distances were constructed for the four botanical varieties,which revealed the existence of different clus-ters. It was concluded that there was abundant intra-variety SSR polymorphism,and with more and more SSR markers being de-veloped,the intrinsic genetic diversity would be detected and the development of genetic map and marker-assisted selection for cultivated peanut would be feasible.

[20] 林茂, 李正强, 郑治洪, 吕建伟, 马天进, 李超, 阚健全 . 贵州省花生地方品种的遗传多样性
作物学报, 2012,38:1387-1396
[本文引用: 1]

Lin M, Li Z Q, Zheng Z H, Lyu J W, Ma T J, Li C, Kan J Q . Genetic diversity of peanut landraces in Guizhou province
Acta Agron Sin, 2012,38:1387-1396 (in Chinese with English abstract)
[本文引用: 1]

[21] 姜慧芳, 任小平, 廖伯寿, 黄家权, 雷永, 陈本银, Guo B Z, Holbrook C C, Upadhyaya H D . 中国花生核心种质的建立及与ICRISAT 花生微核心种质的比较
作物学报, 2008,34:25-30
DOI:10.3724/SP.J.1006.2008.00025URL [本文引用: 1]
fastigiata as well as in plant height and number of total branches than Chinese peanut resource.
Jiang H F, Ren X P, Liao B S, Huang J Q, Lei Y, Chen B Y, Guo B Z, Holbrook C C, Upadhyaya H D . Peanut core collection established in China and compared with ICRISAT mini core collection
Acta Agron Sin, 2008,34:25-30 (in Chinese with English abstract)
DOI:10.3724/SP.J.1006.2008.00025URL [本文引用: 1]
fastigiata as well as in plant height and number of total branches than Chinese peanut resource.

[22] Han Z Q, Gao G Q, Wei P X, Tang R H, Zhong R C . Analysis of DNA polymorphism and genetic relationships in cultivated peanut (Arachis hypogaea L.) using micro-satellite markers
Acta Agron Sin, 2004,30:1097-1101
[本文引用: 1]

[23] 洪彦彬, 梁炫强, 陈小平, 林坤耀, 周桂元, 李少雄, 刘海燕 . 花生栽培种(Arachis hypogaea L.)类型间遗传差异的SSR分析
分子植物育种, 2008,6:71-78
[本文引用: 1]

Hong Y B, Liang X Q, Chen X P, Lin K Y, Zhou G Y, Li S X, Liu H Y . Genetic differences in peanut cultivated types (Arachis hypogaea L.) revealed by SSR polymorphism
Mol Plant Breed, 2008,6:71-78 (in Chinese with English abstract)
[本文引用: 1]

[24] Jiang H F, Liao B S, Ren X P, Lei Y, Mace E, Fu T D, Crouch J H . Comparative assessment of genetic diversity of peanut (Arachis hypogaea L.) genotypes with various levels of resistance to bacteria wilt through SSR and AFLP analyses
J Genet Genomics, 2007,34:544-554
[本文引用: 1]

[25] 黄莉, 任小平, 张晓杰, 陈玉宁, 姜慧芳 . ICRISAT 花生微核心种质农艺性状和黄曲霉抗性关联分析
作物学报, 2012,38:935-946
[本文引用: 1]

Huang L, Ren X P, Zhang X J, Chen Y N, Jiang H F . Association analysis of agronomic traits and resistance to aspergillus flavus in the ICRISAT peanut mini-core collection
Acta Agron Sin, 2012,38:935-946 (in Chinese with English abstract)
[本文引用: 1]

[26] Krishna G K, Zhang J, Burow M, Pittman R N, Delikostadinov S G, Lu Y Z, Puppala N . Genetic diversity analysis in peanut (Arachis hypogaea L.) using microsatellite markers
Cell Mol Biol Lett, 2004,9:685-697
[本文引用: 1]

[27] He G H, Channapatna P . Evaluation of genetic relationships among botanical varieties of cultivated peanut (Arachis hypogaea L.) using AFLP markers
Genet Resour Crop Evol, 2001,48:347-352
[本文引用: 1]

[28] He G H, Meng R H, Gao H, Guo B Z, Gao G Q, Melanie N, Roy N P, Prakash C S . Simple sequence repeat markers for botanical varieties of cultivated peanut (Arachis hypogaea L.)
Euphytica, 2005,142:131-136
[本文引用: 1]

[29] 宗绪晓, 关建平, 王述民, 刘庆昌 . 中国豌豆地方品种SSR标记遗传多样性分析
作物学报, 2008,34:1330-1338
[本文引用: 1]

Zong X X, Guan J P, Wang S M, Liu Q C . Genetic diversity among Chinese pea (Pisum sativum L.) landraces revealed by SSR markers
Acta Agron Sin, 2008,34:1330-1338 (in Chinese with English abstract)
[本文引用: 1]

[30] 宗绪晓, Ford R, Robert R R, 关建平, 王述民 . 豌豆属(Pisum) SSR标记遗传多样性结构鉴别与分析
中国农业科学, 2009,42:36-46
DOI:10.3864/j.issn.0578-1752.2009.01.005URL [本文引用: 1]
【Objective】 Assessing the genetic diversity between wild and cultivated accessions of eight taxonomic groups in two species, five subspecies under Pisum genus, and analyzing the population structure and their genetic relationships among various groups of taxonomy, the study try to verify the fitness of traditionally botanical taxonomic system under Pisum genus and to provide essential information for the exploration and utilization of wild relatives of pea genetic resources. 【Method】 One hundred and ninety-seven Pisum accessions from 62 counties of five continents were employed for SSR analysis using 21 polymorphic primer pairs in this study. Except for cultivated field pea Pisum sativum subsp. sativum var. sativum (94 genotypes), also included were wild relative genotypes that were classified as P. fulvum, P. sativum subsp. abyssinicum, P. sativum subsp. asiaticum, P. sativum subsp. transcaucasicum, P. sativum subsp. elatius var. elatius, P. sativum subsp. elatius var. pumilio and P. sativum subsp. sativum var. arvense (103 genotypes). The PCA analyses and three-dimensional PCA graphs were conducted and drawn by NTSYSpc 2.2d statistical package. Nei78 genetic distances among groups of genetic resources were calculated, and cluster analysis using UPGMA method was carried out by using Popgene V1.32 statistical package, the dendrogram were drawn by MEGA3.1 statistical package. Allelic statistics were carried out by Popgene V1.32. The significance test between groups of genotypes was carried out by Fstat V2.9.3.2 statistical package.【Result】One hundred and four polymorphic bands were amplified using 21 SSR primer pairs with unambiguous unique polymorphic bands. 4.95 alleles were detected by each SSR primer pair in average, of which 65.56% were effective alleles for diversity. PSAD270, PSAC58, PSAA18, PSAC75, PSAA175 and PSAB72 were the most effective SSR pairs. SSR alleles were uniformly distributed among botanical taxon units under pisum genus, but significant difference appeared in most pairwise comparisons for genetic diversity between taxon unit based groups of genetic resources. Genetic diversity level of wild species P. fulvum was much lower than the cultivated species P. sativum. Under species P. sativum, P. sativum ssp. sativum var. sativum and P. sativum ssp. asiaticum were the highest in gentic diversity, followed by P. sativum ssp. elatius var. elatius and P. sativum ssp. transcaucasicum, P. sativum ssp. elatius var. pumilio, P. sativum ssp. sativum var. arvense and P. sativum ssp. abyssinicum were the lowest. Four gene pool clusters were detected under Pisum genus by using PCA analysis. Gene pool “fulvum” mainly consisted of wild species Pisum fulvum, gene pool “abyssinicum” mainly consisted of P. sativum ssp. abyssinicum, and gene pool “arvense” mainly consisted of P. sativum ssp. sativum var. arvense. While gene pool “sativum” were composed by five botanical taxon units, they are P. sativum ssp. asiaticum, P. sativum ssp. elatius var. elatius, P. sativum ssp. transcaucasicum, P. sativum ssp. elatius var. pumilio and P. sativum ssp. sativum var. sativum. “sativum” gene pool constructed the primary gene pool of cultivated genetic resources;“fulvum” gene pool, “abyssinicum” gene pool and “arvense” gene pool together constructed the secondary gene pool of cultivated genetic resources. Pairwise Nei78 genetic distance among botanical taxon based groups of pea genetic resources ranged from 7.531 to 35.956, three large cluster groups were identified based on the UPGMA dendrogram. Group I equals to “sativum” and “arvense” gene pools, Group II equals to “abyssinicum” gene pool, and Group III equals to “fulvum” gene pool. The UPGMA clustering results generally support the PCA clustering results.【Conclusion】 There were significant differences among most botanical groups under Pisum genus, with clear separation of four gene pools for genetic diversity structure. The research results partially support the traditional botanical taxonomy under Pisum genus, and point out its advantage and shortcoming. In order to broaden the genetic bases of pea varieties, the genetic potentials in the four gene pools should be thoroughly exploited.
Zong X X, Ford R, Robert R R, Guan J P, Wang S M . Identification and analysis of genetic diversity structure within Pisum genus based on microsatellite markers
Sci Agric Sin, 2009,42:36-46 (in Chinese with English abstract)
DOI:10.3864/j.issn.0578-1752.2009.01.005URL [本文引用: 1]
【Objective】 Assessing the genetic diversity between wild and cultivated accessions of eight taxonomic groups in two species, five subspecies under Pisum genus, and analyzing the population structure and their genetic relationships among various groups of taxonomy, the study try to verify the fitness of traditionally botanical taxonomic system under Pisum genus and to provide essential information for the exploration and utilization of wild relatives of pea genetic resources. 【Method】 One hundred and ninety-seven Pisum accessions from 62 counties of five continents were employed for SSR analysis using 21 polymorphic primer pairs in this study. Except for cultivated field pea Pisum sativum subsp. sativum var. sativum (94 genotypes), also included were wild relative genotypes that were classified as P. fulvum, P. sativum subsp. abyssinicum, P. sativum subsp. asiaticum, P. sativum subsp. transcaucasicum, P. sativum subsp. elatius var. elatius, P. sativum subsp. elatius var. pumilio and P. sativum subsp. sativum var. arvense (103 genotypes). The PCA analyses and three-dimensional PCA graphs were conducted and drawn by NTSYSpc 2.2d statistical package. Nei78 genetic distances among groups of genetic resources were calculated, and cluster analysis using UPGMA method was carried out by using Popgene V1.32 statistical package, the dendrogram were drawn by MEGA3.1 statistical package. Allelic statistics were carried out by Popgene V1.32. The significance test between groups of genotypes was carried out by Fstat V2.9.3.2 statistical package.【Result】One hundred and four polymorphic bands were amplified using 21 SSR primer pairs with unambiguous unique polymorphic bands. 4.95 alleles were detected by each SSR primer pair in average, of which 65.56% were effective alleles for diversity. PSAD270, PSAC58, PSAA18, PSAC75, PSAA175 and PSAB72 were the most effective SSR pairs. SSR alleles were uniformly distributed among botanical taxon units under pisum genus, but significant difference appeared in most pairwise comparisons for genetic diversity between taxon unit based groups of genetic resources. Genetic diversity level of wild species P. fulvum was much lower than the cultivated species P. sativum. Under species P. sativum, P. sativum ssp. sativum var. sativum and P. sativum ssp. asiaticum were the highest in gentic diversity, followed by P. sativum ssp. elatius var. elatius and P. sativum ssp. transcaucasicum, P. sativum ssp. elatius var. pumilio, P. sativum ssp. sativum var. arvense and P. sativum ssp. abyssinicum were the lowest. Four gene pool clusters were detected under Pisum genus by using PCA analysis. Gene pool “fulvum” mainly consisted of wild species Pisum fulvum, gene pool “abyssinicum” mainly consisted of P. sativum ssp. abyssinicum, and gene pool “arvense” mainly consisted of P. sativum ssp. sativum var. arvense. While gene pool “sativum” were composed by five botanical taxon units, they are P. sativum ssp. asiaticum, P. sativum ssp. elatius var. elatius, P. sativum ssp. transcaucasicum, P. sativum ssp. elatius var. pumilio and P. sativum ssp. sativum var. sativum. “sativum” gene pool constructed the primary gene pool of cultivated genetic resources;“fulvum” gene pool, “abyssinicum” gene pool and “arvense” gene pool together constructed the secondary gene pool of cultivated genetic resources. Pairwise Nei78 genetic distance among botanical taxon based groups of pea genetic resources ranged from 7.531 to 35.956, three large cluster groups were identified based on the UPGMA dendrogram. Group I equals to “sativum” and “arvense” gene pools, Group II equals to “abyssinicum” gene pool, and Group III equals to “fulvum” gene pool. The UPGMA clustering results generally support the PCA clustering results.【Conclusion】 There were significant differences among most botanical groups under Pisum genus, with clear separation of four gene pools for genetic diversity structure. The research results partially support the traditional botanical taxonomy under Pisum genus, and point out its advantage and shortcoming. In order to broaden the genetic bases of pea varieties, the genetic potentials in the four gene pools should be thoroughly exploited.

[31] 刘金, 关建平, 徐东旭, 张晓艳, 顾竟, 宗绪晓 . 小扁豆种质资SSR标记遗传多样性及群体结构分析
作物学报. 2008,34:1901-1909
[本文引用: 1]

Liu J, Guan J P, Xu D X, Zhang X Y, Gu J, Zong X X . Analysis of genetic diversity and population structure in Lentil (Lens culinaris Medik.) germplasm by SSR markers
Acta Agron Sin, 2008,34:1901-1909 (in Chinese with English abstract)
[本文引用: 1]