引种日本多花黑麦草标准品种DUS性状变异分析及应用

doi:10.3864/j.issn.0578-1752.2022.12.015

摘要/Abstract

摘要：

【目的】测定引种自日本的多花黑麦草标准品种DUS测试用性状的一致性和特异性状况,优化我国多花黑麦草DUS测试体系,进而更快速准确地鉴定多花黑麦草新品种(系)。【方法】选择在成都平原种植的7个引自日本的多花黑麦草标准品种,在每个标准品种群体中选取15株长势一致的材料,基于其DUS测试用性状数据对标准品种进行聚类分析,鉴定标准品种在成都平原DUS性状表达情况。通过计算供试材料单株的18个DUS测试用性状的变异系数和方差值,并对测试用性状进行嵌套方差分析,筛选出品种内一致性较高、品种群体间特异性明显的性状。对筛选出来的性状进行概率分级,基于该等级划分为参试国审品种和新品系进行聚类分析和品种鉴定,并对参试品种的测试性状进行相关性分析。【结果】根据标准品种的18个DUS测试用性状的具体数值可以明确地将标准品种聚为7类,且DUS性状表达充分,能较为清晰地相互区分,可以进行后续的相关分析。品种内的一致性分析结果表明,叶色程度、春化后生长习性、孕穗期株幅、孕穗期旗叶宽、孕穗期旗叶长宽比和外颖长在多个品种内部的一致性表现较差。在进行品种群体间特异性分析时发现,孕穗期株高、孕穗期旗叶长宽比和小穗密度在品种群体间的特异性较差,其中孕穗期旗叶长宽比的品种群体间方差分量为54.83%,群体内单株间方差分量为45.17%。对筛选出的10个数量性状进行K-S检验及χ²检验,仅有基部小穗长符合χ²检验。最终将10个数量性状划分为5个等级,基于此性状分级结果对参试的多花黑麦草国审品种及新品系进行鉴定,结果表明供试品种能聚类为8个与品种相互对应的分组,证明了此性状分级对于多花黑麦草品种鉴定的可靠性。此外,基于参试品种性状表达级别绘制的热图结果表明,品种“特高德”在营养生长期的叶长明显区别于其他参试品种,表现为叶长较短;品种“川农2号”表现为春化后株高较高;品种“钻石T×长江2号”的穗下茎节长度明显短于其他参试品种;“达伯瑞”品种群体内一致性表现较差;“川农1号”“川农2号”品种群体内一致性表现较好。同时,筛选出的10个性状之间相关性较低,独立性较强,仅有3组性状间的相关系数大于0.5,可以作为DUS测试用性状。【结论】基于日本标准品种建立的DUS测试用性状分级体系可以应用于我国多花黑麦草的品种鉴定和区分,研究结果为多花黑麦草品种DUS测试方法体系的优化提供了重要的技术和理论参考。

关键词: 多花黑麦草, 数量性状, 概率分级分析, 相关性分析, DUS测试优化

Abstract:

【Objective】This study aimed to determine the consistency and specificity of DUS test traits of Lolium multiflorum (annual ryegrass) standard varieties introduced from Japan, and to optimize the DUS test system of L. multiflorum in China, thus identify new varieties (lines) more quickly and accurately. 【Method】In this study, seven standard varieties of annual ryegrass introduced from Japan were planted in Chengdu Plain, China. Fifteen individuals with the similar growth status were selected from each standard variety. Cluster analysis of the standard varieties was conducted based on their DUS test data to identify the DUS trait expression of the standard varieties in Chengdu Plain. In order to screen out the characters with the high intra-population consistency and obvious inter-population specificity, the coefficient of variation and variance values of 18 tested DUS characters were calculated, and the nested variance analysis of the studied characters was also conducted. Furthermore, the probability classification was conducted based on the selected characters, which was used to clustering analysis and identification of the tested national approved varieties and new lines, and to evaluate the correlation coefficients of test traits. 【Result】According to the values of 18 DUS test traits of standard varieties, the standard varieties could be clearly clustered into 7 groups. The DUS traits were fully expressed and could be clearly distinguished from each other, which were suitable for subsequent correlation analysis. The results of intraspecific consistency analysis showed that, six traits, including leaf color degree, growth habit after vernalization, plant width at booting stage, flag leaf width at booting stage, flag leaf length-width ratio at booting stage, and spikelet length, possessed the poor intra-population consistency for multiple varieties. When conducting the inter-population specificity analysis of varieties, however, the poor specificity among the tested varieties was found in plant height at booting stage, ratio of flag leaf length to width at booting stage and spikelet density among cultivars, among which the variance component between populations of flag leaf length-width ratio at booting stage was 54.83%, and the variance component between individuals within populations was 45.17%. The selected ten quantitative traits were conducted the K-S test and χ² test analysis, however, only the basal spikelet length conformed to χ² test. Afterwards, the ten quantitative traits were divided into five grades. On the basis of these traits grading criterion, the identification of national approved varieties and new lines of L. multiflorum. The results showed that the tested varieties and new lines of annual ryegrass could be clustered into eight groups according to its classification results and corresponding to varieties origin, which revealed the reliability of traits grading criterion built in this study on variety identification. The heat map drawn based on the trait values of the tested varieties showed that the leaf length of ‘Tetragold’ was significantly different from that of other tested varieties in the vegetative growth period, which was manifested as shorter leaf length; the variety ‘Chuannong No. 2’ showed higher plant height after vernalization; the internode length under spike of ‘Diamond T×changjiang No. 2’ was significantly shorter than that of other varieties; the poor intra-population consistency of ‘Double Barrel’, ‘Chuannong No. 1’ and ‘Chuannong No. 2’ showed good intra-population consistency. The low correction and strong independence were existed within the selected ten traits, indicating that these traits were very well suited for DUS test. 【Conclusion】The character classification system for DUS test based on Japanese standard varieties could be applied to the identification and distinguish of L. multiflorum varieties released in China. This study could provide the important technical and theoretical reference for the optimization of DUS test method system of annual ryegrass varieties.

Key words: Lolium multiflorum, quantitative characters, probability classification analysis, correlation analysis, DUS test optimization

冯俊杰,赵文达,张新全,刘英杰,袁帅,董志晓,熊毅,熊艳丽,凌瑶,马啸. 引种日本多花黑麦草标准品种DUS性状变异分析及应用[J]. 中国农业科学, 2022, 55(12): 2447-2460.

FENG JunJie,ZHAO WenDa,ZHANG XinQuan,LIU YingJie,YUAN Shuai,DONG ZhiXiao,XIONG Yi,XIONG YanLi,LING Yao,MA Xiao. DUS Traits Variation Analysis and Application of Standard Varieties of Lolium multiflorum Introduced from Japan[J]. Scientia Agricultura Sinica, 2022, 55(12): 2447-2460.

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链接本文: https://www.chinaagrisci.com/CN/10.3864/j.issn.0578-1752.2022.12.015

https://www.chinaagrisci.com/CN/Y2022/V55/I12/2447

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编号 No.	品种名称 Variety name	登记时间 Registration time	选育单位 Breeding organization	品种类型 Type
1	Satiaoba	——	日本 Japan	日本标准品种 Japanese standard varieties
2	Yuosyunn	——	日本 Japan	日本标准品种 Japanese standard varieties
3	Nagahahikari	——	日本 Japan	日本标准品种 Japanese standard varieties
4	Waseaoba	——	日本 Japan	日本标准品种 Japanese standard varieties
5	Hitatihikari	——	日本 Japan	日本标准品种 Japanese standard varieties
6	Shiwasuaoba	——	日本 Japan	日本标准品种 Japanese standard varieties
7	Akiaoba	——	日本 Japan	日本标准品种 Japanese standard varieties
8	特高德Tetragold	2001	广东省饲草饲料站 Guangdong Feeding Station	国审引进品种 Imported varieties of national audit
9	剑宝Jumbo	2015	四川省畜牧科学研究院 Sichuan Academy of Animal Science	国审引进品种 Imported varieties of national audit
10	川农1号Chuannong No. 1	2016	四川农业大学 Sichuan Agricultural University	国审育成品种Varieties bred varieties
11	达伯瑞Double Barrel	2012	云南省草山饲料工作站 Yunnan Grassy Hill Feed Workstation	国审引进品种 Imported varieties of national audit
12	邦德Abundant	2008	云南省草山饲料工作站 Yunnan Grassy Hill Feed Workstation	国审引进品种 Imported varieties of national audit
13	南黑1号Nanhei No. 1	——	四川省农科院蚕桑所 Sericulture Institute, Sichuan Academy of Agricultural Sciences	新品系 New strain
14	川农2号Chuannong No. 2	——	四川农业大学 Sichuan Agricultural University	新品系 New strain
15	钻石T×长江2号 Diamond T×changjiang No. 2	——	四川农业大学 Sichuan Agricultural University	新品系 New strain

代码 Code	名称 Name	描述 Description	代码 Code	名称 Name	描述 Description
00	发芽期 Germination stage	干种子 Dry seeds	41	孕穗期 Boot stage	旗叶叶鞘伸展 Flag leaf sheath extension
01		开始吸水 Start absorbing water	45		穗苞膨大 Bracts swelled
03		吸水结束 End of water absorption	47		第1叶鞘张开 1st leaf sheath open
05		胚根从颖果中露出 Embryo roots exposed from glume	49		第1芒可见 First awn visible
07		胚芽鞘从颖果中露出 Germ sheath emerged from glume	50	花序形成期 Inflorescence formation stage	花序第1小穗可见 Inflorescences 1st spikelet visible
09		叶刚露出胚芽鞘顶端 Leaf just exposed the top of germ sheath	52		1/4花序出现 1 / 4 inflorescence appeared
10	幼苗生长期 Seedling season of growth	第1叶从胚芽鞘长出 The first leaf grows from the germ sheath	54		1/2花序出现 1 / 2 inflorescence appeared
15		第1叶展开 The first leaf unfolds	56		3/4花序出现 3 / 4 inflorescence appeared
19		第5叶或者更多叶展开 5th or more leaves unfold	58		整个花序出现 The whole inflorescence appears
20	分蘖期 Tillering stage	仅有主茎 Main stem only	60	开花期 Flowering period	开花开始 Onset of flowering
23		主茎和3个分蘖 Main stem and 3 tillers	64		开花一半 Flowering half
25		主茎和5个分蘖 Main stem and 5 tillers	68		开花结束 Flowering end
29		主茎和9个以上分蘖 Main stem and more than 9 tillers	70	成熟期 Maturity	颖果结束 The end of caryopsis
30	拔节期 Jointing stage	假茎直立 Pseudostem erect	72		种子休眠 Seed dormancy
31		第1节出现 Section 1 appears	76		种子休眠结束 End of seed dormancy
35		第5节出现 Section 5 appears	78		诱发第二次休眠 Inducing second sleep
39		旗叶可见 Flag leaf visible	80		二次休眠消失 Secondary sleep disappears

编号 Number	性状 Characteristics	观测时期对应代码 Corresponding code for Survey period	编号 Number	性状 Characteristics	观测时期对应代码 Corresponding code for Survey period
C1	叶长（营养生长期） Leaf: length(at vegetative stage)	20-29	C10	孕穗期旗叶宽 Flag leaf: width at inflorescence emergence	50
C2	叶宽（营养生长期） Leaf: width(at vegetative stage)	20-29	C11	旗叶长宽比 Flag leaf: length/width ratio	50
C3	叶色（营养生长期） Leaf: intensity of green color (at vegetative stage)	20-29	C12	最长茎秆：长度（包括花序） Plant: length of longest stem, inflorescence included (when fully expanded)	60-68
C4	株高（春化后） Leaf: intensity of green color	30-39	C13	穗下茎节长度 Plant: length of upper internode	60-68
C5	生长习性（春化后） Plant: vegetative growth habit (after vernalization)	30-39	C14	花序长 Inflorescence: length	60-68
C6	孕穗期（春化后） Plant: time of inflorescence emergence (after vernalization)	50	C15	小穗数 Inflorescence: number of spikelet	60-68
C7	株高（孕穗期） Plant: natural height at inflorescence emergence	50	C16	小穗密度（每个小穗着生长度） Inflorescence: density ( dividing characteristic 16 by characteristic 17)	60-68
C8	株幅（孕穗期） Plant: width at inflorescence emergence	50	C17	外颖长 Inflorescence: length of outer glume on basal spikelet	60-68
C9	旗叶长（孕穗期） Flag leaf: length at inflorescence emergence	50	C18	基部小穗长（不包括芒） Inflorescence: length of basal spikelet excluding awn	60-68

方差分量 Variance component	品种群体间（%） Between varietal groups	群体内单株间（%） Per plant of varieties	误差 Error	显著性 Significance
C1	89.41	5.41	5.07	**
C2	92.16	2.16	5.68	**
C3	90.87	9.13	—	**
C4	77.47	22.53	—	**
C5	90.66	9.34	—	**
C6	97.49	2.51	—	**
C7	54.83	45.17	—	**
C8	80.65	19.35	—	**
C9	88.15	6.07	5.78	**
C10	90.29	4.93	4.78	**
C11	59.26	10.67	30.06	**
C12	87.54	9.36	3.1	**
C13	90.40	4.07	5.53	**
C14	93.69	1.34	4.97	**
C15	94.75	1.08	4.17	**
C16	66.93	2.9	30.16	**
C17	88.92	6.02	5.06	**
C18	96.70	2.51	0.79	**

性状 Characteristics	正极差 Positive	负极差 Negative	K-S值 K-S value	Sig值 Sig value	χ²-P值 χ²-P value
C1	0.040	-0.022	0.040	0.200	1.000
C2	0.024	-0.031	0.031	0.200	1.000
C4	0.085	-0.040	0.085	0.058	1.000
C6	0.059	-0.084	0.084	0.067	0.560
C9	0.026	-0.020	0.026	0.200	1.000
C12	0.034	-0.044	0.044	0.200	1.000
C13	0.033	-0.029	0.033	0.200	1.000
C14	0.027	-0.033	0.033	0.200	1.000
C15	0.074	-0.050	0.074	0.058	0.000
C18	0.123	-0.112	0.123	0.000	1.000