崔国庆¹, 王世明¹, 马福盈¹, 汪会¹, 向朝中², 李云峰¹, 何光华¹, 张长伟¹, 杨正林¹, 凌英华¹, 赵芳明^,1,*1 西南大学水稻研究所/西南大学农业科学研究院, 重庆 400715
2 重庆市丰都县龙孔镇农业服务中心, 重庆408200

Identification of Rice Chromosome Segment Substitution Line Z1377 with Increased Plant Height and QTL Mapping for Agronomic Important Traits

CUI Guo-Qing¹, WANG Shi-Ming¹, MA Fu-Ying¹, WANG Hui¹, XIANG Chao-Zhong², LI Yun-Feng¹, HE Guang-Hua¹, ZHANG Chang-Wei¹, YANG Zheng-Lin¹, LING Ying-Hua¹, ZHAO Fang-Ming^,1,* 1 Rice Research Institute, Southwest University / Academy of Agricultural Sciences, Southwest University, Chongqing 400715, China
2 Center of Agricultural Serve at Longkong Town, Fengdu County, Chongqing 408200, China

通讯作者: ^* 通信作者(Corresponding author): 赵芳明, E-mail: zhaofangming2004@163.com

第一联系人: E-mail: 271014808@qq.com
收稿日期:2018-01-5接受日期:2018-06-12网络出版日期:2018-06-20

基金资助:

本研究由国家重点研发计划项目.2017YFD0100202 重庆市科委主题专项.CSTC, 2016shms-ztzx0017 西南大学基本业务费专项创新团队项目.XDJK2017A004

Received:2018-01-5Accepted:2018-06-12Online:2018-06-20

Fund supported:

This study was supported by the National Key Research and Development Program of China.2017YFD0100202 the Project of Chongqing Science & Technology Commission.CSTC, 2016shms-ztzx0017 the Fundamental Research Funds for the Central Universities.XDJK2017A004

摘要
株高是水稻重要的农艺性状, 往往与产量相关性状密切关联, 在水稻育种中有重要利用价值。本研究以日本晴为受体、缙恢35为供体亲本, 经表型和分子标记双重选择, 鉴定了一个水稻高秆染色体片段代换系Z1377。Z1377共含有18个代换片段, 平均代换长度为2.95 Mb。与日本晴相比, Z1377的株高、倒一节间至倒四节间长、穗长、一次枝梗数、二次枝梗数、粒长、每穗实粒数、总粒数显著增加; 粒宽显著变细, 有效穗数、结实率显著减少, 但仍达86.75%。用日本晴与Z1377杂交构建的次级F₂群体共检测到16个相关QTL, 分布于第2、第3、第4、第5、第6、第7和第9染色体。其中有8个可能与已克隆基因等位, 如GW2、EUI1、ZFP185等, 另8个如qPH3等尚未见报道。Z1377的株高由一个主效QTL (qPH3)和一个微效QTL (qPH5)控制, 其中qPH3的贡献率达28.59%。而且, 在F₂群体中, 高秆和矮秆基本呈现双峰分布, 经卡平方测验, 符合3∶1分离比, 表明高秆对矮秆显性, 并主要由qPH3负责。这将为该主效基因的精细定位和克隆奠定基础, 同时为进一步选育含2~3个代换片段的中高优良染色体片段代换系并应用于育种奠定基础。
关键词： 水稻;染色体片段代换系;株高;产量相关性状;QTL定位

Abstract
Plant height is an important agronomic trait in rice, usually relating to yield-related traits. Here, a novel rice chromosome segment substitution line Z1377 with increased plant height was identified from recipient Nipponbare and donor Jinhui 35 through selection of both phenotype and molecular marker. Z1377 carried 18 substitution segments with average substitution length of 2.95 Mb. Compared with Nipponbare, Z1377 had significantly increased plant height, length of the 1st, 2nd, 3rd, and 4th internode, panicle length, number of primary and secondary branches and grain length, and decreased grain width, number of panicles per plant and seed setting ratio. However, the seed setting ratio was still 86.75%. Furthermore, F₂ population from crosses between Nipponbare and Z1377 was used to map QTLs for plant height and other important agronomic traits. A total of 16 QTLs were detected, of which eight had been reported with the cloned genes such as GW2, EUI1, ZFP185, and the other eight were still not reported, such as qPH3. The plant height of Z1377 was mainly controlled by a major QTL (qPH3) with the explained phenotypic variance of 28.59% and a minor QTL (qPH5). Moreover, the high and dwarf plants basically displayed a bimodal distribution in the F₂ population, and fitted to 3:1 segregation ratio by Chi-square test, indicating that high plant is dominant to dwarf plants and mainly conferred by qPH3. These results lay a foundation for fine mapping and cloning qPH3, meanwhile, also provide good bases for developing excellent chromosome segment substitution lines with moderate height plant carrying 2-3 substitution segments to be used in breeding.
Keywords：rice;chromosome segment substitution lines;plant height;yield-related traits;QTL mapping


PDF (1263KB)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
崔国庆, 王世明, 马福盈, 汪会, 向朝中, 李云峰, 何光华, 张长伟, 杨正林, 凌英华, 赵芳明. 水稻高秆染色体片段代换系Z1377的鉴定及重要农艺性状QTL定位[J]. 作物学报, 2018, 44(10): 1477-1484. doi:10.3724/SP.J.1006.2018.01477
CUI Guo-Qing, WANG Shi-Ming, MA Fu-Ying, WANG Hui, XIANG Chao-Zhong, LI Yun-Feng, HE Guang-Hua, ZHANG Chang-Wei, YANG Zheng-Lin, LING Ying-Hua, ZHAO Fang-Ming. Identification of Rice Chromosome Segment Substitution Line Z1377 with Increased Plant Height and QTL Mapping for Agronomic Important Traits[J]. Acta Agronomica Sinica, 2018, 44(10): 1477-1484. doi:10.3724/SP.J.1006.2018.01477


自20世纪50年代的水稻“第一次绿色革命”以来, 水稻产量得到大幅提高, 该技术的关键是通过株型矮化以提高收获指数来获取高产。然而, 水稻收获指数目前已经很高, 通过该途径提高产量已较难, 矮秆已成为限制水稻生物量的关键因素。随着科技的高速发展, 通过发展半高秆抗倒品种提高产量已成为重要途径。因此深入挖掘水稻株高发育相关基因, 阐明水稻株高形成的分子机制, 对于水稻株型育种有重要意义。

目前通过克隆水稻株高相关基因, 初步揭示了水稻株高主要由赤霉素(gibberellin, GA)、油菜素内酯(brassionosteriod, BR)和独角金内酯(strigolactones, SL) 3种水稻内源激素调控^[1]。“绿色革命”基因sd1编码GA合成途径中的GA20氧化酶^[2]。多个调控GA合成途径的基因如D18^[3]、D35^[4]等突变, 使该途径发生障碍, 导致不同程度的水稻矮化。BR是控制水稻株高的第2种关键激素, 能促进茎的伸长。矮秆突变体brd1株高只有10~30 cm, 由OsBR6ox控制, 是水稻BR生物合成的一个关键酶, 即BR-6氧化酶, 属于细胞色素P450家族^[5]; 矮秆基因 D11的编码蛋白CYP724B1, 也属于P450家族, 通过外源施加BR后可以恢复表型^[6]。还有一些基因, 如DWARF AND LOW-TILLERING, 突变后外源施加BR无法恢复表型, 主要通过调控BR传导途径来影响水稻株高^[7]。此外, 独角金内酯(SL)也参与调控水稻株高, 如d10编码类胡萝卜素裂解双加氧酶OsCCD8, 该酶参与SL的生物合成^[8]。

株高是遗传复杂的农艺性状, 涉及到大量的基因, 而且往往与产量性状相关联。因而, 目前克隆基因仍不能完全阐明株高发育的分子机制, 有必要发掘新的株高相关基因。尤其, 目前发现的株高突变体, 多数均表现矮秆、穗型和谷粒偏小, 如d18^[3]、brd1^[5]、d11^[6]等, 较难直接用于聚合育种。本研究以日本晴为受体亲本、缙恢35为供体亲本鉴定了一个18代换片段的水稻高秆染色体片段代换系Z1377, 全面分析该代换系并进行株高等重要农艺性状QTL定位, 以期为克隆株高等性状的主效QTL奠定基础。

1 材料与方法

1.1 试验材料

18代换片段的水稻染色体片段代换系Z1377以日本晴为受体、缙恢35为供体亲本, 经连续回交自交结合表型和分子标记双向选择选育而成。缙恢35是西南大学选育的优良三系恢复系, 选择其为供体亲本是为鉴定其在测序品种日本晴的遗传背景下代换片段上的优良等位基因。QTL定位群体材料为日本晴和Z1377杂交构建的次级F₂群体。

1.2 试验方法

1.2.1 Z1377代换片段鉴定 使用均匀分布于水稻全基因组的429个标记, 进行日本晴和缙恢35的多态性分析, 选出253个多态性标记^[9], 然后用这些多态性标记从BC₂F₁进行分子标记和表型双向选择, 直至BC₂F₁₀初步选育成18代换片段的Z1377。参照向佳等^[10]描述的方法鉴定代换片段, 当某一标记的检测结果与受体亲本的带型(A)一致时, 认为该段DNA来自日本晴基因组; 当某一标记的检测结果与供体亲本带型(B)一致时, 则认为该段DNA来自缙恢35基因组。连续B标记间所在区段表示代换片段。按Paterson等^[11]方法计算代换片段长度。

1.2.2 材料种植方法 2016年以日本晴与Z1377杂交, 收获F₁并于同年秋季在海南种植, 并收获F₂种子。2017年3月8日, 用亲本(日本晴和Z1377)及F₂群体种子于重庆西南大学水稻研究所实验基地育苗。4月15日, 以株、行距分别为16.67 cm和26.67 cm移栽日本晴和Z1377各30株及88个F₂单株于同一试验田, 常规管理。

1.2.3 农艺性状调查 Z1377与日本晴的抽穗期相当, 仅晚4 d, 因而QTL定位时没有考察该性状。在成熟期, 平地面收取日本晴和Z1377各10株以及F₂群体88株作为样本, 考察株高、有效穗数、各节间长度、穗长、一次枝梗数、二次枝梗数、粒长、粒宽、实粒数、总粒数、千粒重和单株产量。其中, 株高和各节间长度测定每株主穗的长度。穗长、一次枝梗数、二次枝梗数、每穗实粒数和总粒数测定用每株全部有效穗的平均值。千粒重测定, 以日本晴和Z1377各取饱满均匀的3000粒籽粒, 每1000粒作为一个单元在电子天平称重, 重复3次; F₂群体每株以200粒作为一个单元称重, 乘以5, 重复3次, 计平均数。以每穗实粒数占每穗总粒数的百分比表示结实率。以粒长与粒宽的比为谷粒长宽比。Microsoft Excel 2003统计10株日本晴和Z1377的上述性状的平均值及F₂群体各参数如平均数、最小值、最大值、偏度和峰度, 并进行t测验。

1.2.4 分子定位 采用CTAB法提取亲本及用于QTL定位的88个F₂单株的DNA。参照向佳等^[10]描述的PCR扩增和非变性聚丙烯酰胺凝胶电泳和快速银染的方法。将日本晴带型标以“-1”, Z1377带型标以“1”, 双亲带型标以“0”, 缺失带型标以“.”。将F₂的88个单株的各性状平均值及标记赋予值一同用于QTL定位。在SAS统计软件上, 利用加州大学河滨分校徐世忠教授编写的HPMIXED程序限制性最大似然(REML)法(略加修改)定位QTL ^[12], 以P < 0.05为阈值决定一个QTL是否与代换片段上的某标记连锁。

2 结果与分析

2.1 Z1377的代换片段鉴定

用Z1377代换片段上的56个SSR标记及代换片段外的24个SSR标记对10株Z1377进行进一步代换片段鉴定和遗传背景纯度检测, 发现10个单株的代换片段一致, 且没有检测出除代换片段外的缙恢35其他残留片段, 说明Z1377已纯合。Z1377共含有来自缙恢35的18个代换片段, 分布于水稻12条染色体(图1)。总代换长度为53.1 Mb, 最长代换长度为8.2 Mb, 最短代换长度为0.7 Mb, 平均代换长度为2.95 Mb。

图1

新窗口打开|下载原图ZIP|生成PPT
图1Z1377的代换片段

每条染色体左侧为物理距离(Mb)和定位的QTL; 右侧为标记名称、代换区间(框内标记)和代换长度(黑箭头指向)。
Fig. 1Substitution segments of Z1377

Physical distances (Mb) and mapped QTL are marked at the left of each chromosome; Markers, substitution interval squared by frame, and substitution length (black arrow direction) are displayed at the right of each chromosome.

2.2 Z1377的农艺性状分析

与受体日本晴相比, Z1377的株高极显著增加(图2和表1)。Z1377除比日本晴多了2个节间(倒五节和倒六节)外, 穗长和其他4个节间长度均显著增加。说明Z1377的株高增加是节间数目和节间长度及穗长共同增加的结果。此外, Z1377的一次枝梗数、二次枝梗数、粒长、实粒数、总粒数显著增加; 有效穗数、粒宽、结实率显著减小, 然而, 其结实率仍达86.75%。Z1377的千粒重和单株重与日本晴无显著差异(表1)。暗示了Z1377中携带了来自缙恢35的多个差异性状的QTL。

Table 1
表1
表1成熟期的日本晴和Z1377及F₂群体各性状统计参数
Table 1Statistical parameters of different traits in Nipponbare, Z1377, and the F₂ population

性状 Trait	平均值±标准差(亲本) Mean±SD (parents)		F2 群体 F2 population
	日本晴 Nipponbare	Z1377	平均值±标准差 Mean±SD	范围 Range	偏度 Skewness	峰度 Kurtosis
株高Plant height (cm)	89.68±2.65**	171.39±3.87**	130.07±21.39	81.60-169.50	-0.62	-0.58
有效穗数Panicle number	13.65±3.30**	5.20±1.17**	8.70±4.17	1.00-27.00	1.24	3.97
倒一节Length of the first internode (cm)	39.02±2.16**	56.02±1.81**	45.69±6.23	32.00-59.80	-0.17	-0.65
倒二节Length of the second internode (cm)	18.32±1.37**	26.58±1.27**	22.45±4.71	10.50-32.30	-0.36	-0.03
倒三节Length of the third internode (cm)	10.19±1.67**	22.45±1.64**	15.96±4.60	3.80-26.20	-0.21	-0.12
倒四节Length of the fourth internode (cm)	1.84±0.75**	17.46±1.05**	11.96±3.54	1.80-19.50	-0.89	1.05
倒五节Length of the fifth internode (cm)	—	11.08±2.78	7.03±3.20	1.70-13.70	0.20	-0.88
倒六节Length of the sixth internode (cm)	—	4.00±1.29	5.21±3.00	1.30-10.10	0.15	-1.52
穗长Panicle length (cm)	20.09±2.17**	33.49±3.17**	27.86±5.39	18.00-42.77	0.22	-0.32
一次枝梗数Number of primary branch	8.43±1.73**	15.33±1.91**	12.12±2.63	7.21-18.10	-0.03	-0.56
二次枝梗数Number of secondary branch	26.47±7.51**	71.69±22.46**	49.85±17.90	21.13-98.90	0.32	-0.29
粒长Grain length (mm)	7.01±0.10**	10.02±0.16**	8.45±0.75	7.00-9.87	-0.35	-0.85
粒宽Grain width (mm)	3.42±0.07**	2.92±0.08**	3.14±0.25	2.50-3.60	-0.41	-0.62
实粒数Grains per panicle	96.99±8.62**	233.06±55.96**	91.69±53.23	2.00-224.00	0.28	-0.44
总粒数Spikelets per panicle	103.04±8.82**	272.65±69.96**	187.18±71.76	42.00-370.00	0.39	-0.25
结实率Seed setting ratio (%)	94.13±0.01**	86.75±2.97**	0.54±0.31	0.02-0.95	-0.29	-1.42
千粒重1000-grain weight (g)	25.60±0.56	25.37±0.66	24.61±3.21	16.60-31.40	-0.26	-0.44
单株产量Yield per plant (g)	33.82±7.15	30.77±7.43	22.42±17.71	0.27-82.4	1.16	1.62

^* and ^** indicate significant difference between traits at P < 0.05 and P < 0.01, respectively.
^* 和 ^**分别表示性状间存在显著(P < 0.05)或极显著(P < 0.01)差异。

新窗口打开|下载CSV

图2

新窗口打开|下载原图ZIP|生成PPT
图2日本晴和Z1377的表型

Fig. 2Phenotype of Nipponbare and Z1377

2.3 Z1377的株高等性状在F₂群体的分布和遗传分析

以日本晴为母本与高秆代换系Z1377杂交, F₁植株的株高偏高, 说明增加株高可能为显性遗传。在88株的F₂群体中, 株高呈负偏态双峰分布, 矮株峰值主要集中在80~110 cm, 合计18株; 高株峰值主要集中在110~170 cm, 合计70株。经卡平方测验, 高株(70)∶矮株(18)符合3∶1分离比(χ² = 0.74 < χ²_(0.05,1) = 3.84)(图3), 表明Z1377的株高增加主要受主效QTL显性控制。其他的多数性状基本呈正态分布, -0.5≤偏度≤0.5; -0.5≤峰度≤2.0, 如倒一至倒五节间长、穗长、一次枝梗数、二次枝梗数、每穗实粒数、总粒数及千粒重。而有效穗数呈正偏态且峰度很高, 倒六节间长和结实率图形接近半矩形; 单株产量呈较强正偏态(图3)。

图3

新窗口打开|下载原图ZIP|生成PPT
图3日本晴与Z1377杂交的F₂群体株高分布

Fig. 3Distribution of plant height in F₂ population from the cross between Nipponbare and Z1377

2.4 Z1377重要农艺性状QTL定位

共鉴定出16个重要农艺性状QTL, 分布于第2、第3、第4、第5、第6、第7、第9染色体。影响株高的QTL共有2个, 分别为qPH3和qPH5, 位于第3和第5染色体, 来自缙恢35的等位基因分别可增加7.52 cm和4.65 cm的株高, 贡献率分别为28.59%和10.96%, 其中qPH3是一个主效株高QTL。倒一节间长qLFI2, 位于第2染色体, 估计可增加长度为1.15 cm, 贡献率为3.65%。倒三节长QTL共3个, 其中qLTI4位于第4染色体, 估计可增加长度为1.19 cm, 贡献率为8.50%; qLTI6-1和qLTI6-2均位于第6染色体, 估计增加长度分别为1.80 cm和1.13 cm, 贡献率分别为19.41%和7.59%。倒四节长qLFOI3, 位于第3染色体, 估计增加长度为1.31 cm, 贡献率为16.44%。穗长qPL2、一次枝梗数qNPB2、二次枝梗数qNSB2、粒宽qGW2、总粒数qSPP2, 均位于第2染色体, 与RM6378连锁, 估计效应分别为1.30 cm、0.54、4.57、-0.06 mm和14.68, 贡献率分别为11.00%、8.09%、11.09%、7.43%和5.82%。粒长qGL3, 位于第3染色体, 估计增加粒长为0.18, 贡献率为12.38%。2个实粒数QTL qGPP6和qGPP9, 分别位于第6和第9染色体, 估计可分别增加每穗38.86粒和90.26粒, 贡献率分别为0.50%和2.71%。有效穗qPN7位于第7染色体, 暗示来自缙恢35的等位基因估计可减少每株2.53个有效穗, 贡献率为3.9%。

Table 2
表2
表2水稻重要农艺性状QTL
Table 2QTLs for agronomic important traits in rice

性状 Trait	QTL	染色体 Chr.	连锁标记 Linked marker	估计效应 Estimated effect	贡献率 Var. (%)	P值 P-value
株高Plant height (cm)	qPH3	3	RM14412	7.52	28.59	0.0008
株高Plant height (cm)	qPH5	5	RM3321	4.65	10.96	0.0305
倒一节Length of the first internode (cm)	qLFI2	2	RM6378	1.15	3.65	0.0474
倒三节Length of the third internode (cm)	qLTI4	4	RM7187	1.19	8.50	0.0367
倒三节长Length of the third internode (cm)	qLTI6-1	6	RM3183	1.80	19.41	0.0031
倒三节长Length of the third internode (cm)	qLTI6-2	6	RM103	1.13	7.59	0.0484
倒四节长Length of the fourth internode (cm)	qLFOI3	3	RM14412	1.31	16.44	0.0043
穗长Panicle length (cm)	qPL2	2	RM6378	1.30	11.00	0.0124
一次枝梗数Number of primary branch	qNPB2	2	RM6378	0.54	8.09	0.0188
二次枝梗数Number of secondary branch	qNSB2	2	RM6378	4.57	11.09	0.0082
粒长Grain length (mm)	qGL3	3	RM14412	0.18	12.38	0.0220
粒长Grain width (mm)	qGW2	2	RM6378	-0.06	7.43	0.0210
实粒数Grains per panicle	qGPP6	6	RM3183	38.86	0.50	0.0062
实粒数Grains per panicle	qGPP9	9	RM7048	90.26	2.71	0.0025
总粒数Spikelets per panicle	qSPP2	2	RM6378	14.68	5.82	0.0433
有效穗数Panicle number	qPN7	7	RM5481	-2.53	3.90	0.0293

新窗口打开|下载CSV

3 讨论

利用染色体片段代换系可简化复杂遗传性状的研究, 并有利于农艺性状基因的鉴定^[13]。本研究鉴定了一个水稻高秆染色体片段代换系Z1377, 其株高的增加由其节间的增多(增加2节)以及各节间和穗长的共同增加所致。然而, 其抽穗期与日本晴相当, 仅晚4 d, 表明Z1377的生长速率快于日本晴, 与抽穗期延长的株高增加^[10]相比, 具有更重要的研究价值。目前现有水稻品种的收获指数已接近植物生理学家认为的上限0.6, 进一步高产必须首先在生物量上有所突破, 而其重要途径之一就是增加株高^[14]。尽管Z1377在育种上直接应用嫌太高(株高171.39 cm), 但可在明确其株高及重要性状的遗传行为基础上进一步将其分解加以利用。

为了进一步明确Z1377株高等差异性状的遗传行为, 本研究利用日本晴与Z1377杂交产生的次级F₂群体鉴定出了16个重要农艺性状的QTL, 除粒宽和结实率QTL外, 其他都是正效应。2个株高QTL (qPH3和qPH5)的遗传效应分别增加株高7.52 cm和4.52 cm, 贡献率分别为28.59%和10.96%。说明Z1377的株高遗传为一个主基因加一个微效基因模式。然而, 在F₂群体中, 高秆和矮秆基本呈现双峰分布, 经卡平方测验符合3∶1分离比, 表明高秆对矮秆显性, 主要由qPH3负责。经与已定位的QTL比较, qPH3是一个尚未报道的株高QTL, 可能是一个新基因。而qPH5, 与已经克隆的水稻长穗颈基因EUI1位于同一区间, 该基因编码细胞色素P450单加氧酶CYP714D1, eui1控制的隐性高秆突变体通过倒一节显著伸长增加株高^[15]。穗长基因qPL2 与已克隆的影响穗长基因ZFP185^[16]及调控水稻穗结构基因FUWA在相同的区间, 其中ZFP185含有2个保守锌指结构域, 但缺乏转录激活能力, 过表达ZFP185植株表现半矮化、短节间和穗; FUWA编码包含NHL结构域的蛋白, fuwa突变体株高略矮, 分蘖数减少, 穗变短, 二次枝梗减少, 每穗粒数减少, 籽粒变宽、变厚、变短, 千粒重增加^[17]。而本研究鉴定的倒一节间长qLFI2、一、二次枝梗数qNPB2和qNSB2以及窄粒qGW2和多粒数qSPP2也均在该区间。此外, Li等^[18]也在此区间(RM3732-RM1358)报道了一次枝梗数目增多的大穗基因(LARGER PANICLE, LP), 可能与本研究发现的qNPB2是等位基因。Song等^[19]同样在该区间克隆了一个控制水稻粒宽的QTL-GW2, 可能与本研究中qGW2等位, GW2编码一个环型E3泛素连接酶。这些基因之间有着怎么样的调控关系有待进一步研究, 说明Z1377是研究这些基因关系的重要材料。本研究鉴定的粒长基因qGL3与增加籽粒长度和重量的PGL1^[20]在同一座位(RM1332-RM175)。本研究鉴定的多实粒数基因qGPP9与增加粒数的等位基因DN1^[21]和DEP1^[22]在相同的区间(RM5657- RM6491)。表明这些基因/QTL比较稳定, 能在不同遗传背景和环境中被检出。本研究中的qPH3、qLFI2、qLTI4、qLTI6-1、qLTI6-2、qLFOI3、qGPP6、qPN7均未见报道, 可能是新的位点。因而, 我们一方面可进一步精细定位和克隆主效株高QTL-qPH3, 为qPH3调控株高形成的分子机制研究奠定基础; 另一方面, 可据F₂各株系的QTL定位结果和株高等表型结果深入分析, 如在表型上选择中高且其他性状优良, 在分子标记上将qPH5、qLFI2、qLTI4、qLTI6-1、qLTI6-2等QTL的标记选为日本晴背景, 将这些增加株高的QTL去除, 而保留多粒数qGPP9等有利基因, 就可选出含2~3个代换片段的中高优良染色体片段代换系, 进而在育种中加以利用。

株高QTL常与产量相关性状QTL成簇状分布, 或一因多效, 或紧密连锁。本研究检出的株高qPH3、倒四节间长qLFOI3和粒长qGL3同连锁于第3染色体RM14412。倒一节间长qLFI2、一、二次枝梗数qNPB2和qNSB2以及窄粒qGW2和多粒数qSPP2也同连锁于第2染色体RM6378标记。刘胜男等也在同一代换片段上检测出了株高qPH8、粒长qGS8和千粒重qKGW8^[23]。Ghd7也是一个一因多效的基因, 同时控制水稻株高、每穗粒数和抽穗期^[24]。这些相关性状QTL成簇状分布的现象相当普遍, 被称为“多因子连锁(linked factors)”^[25], 可能是自然选择中性状的有利协同进化的结果^[26]。在聚合育种中, 当目标性状QTL与有利性状连锁时, 便于利用; 然而往往是不利性状QTL连锁较多, 这就需要我们打破连锁累赘, 才能达到预期目标。

4 结论

鉴定了一个以日本晴为受体、缙恢35为供体亲本的水稻高秆染色体片段代换系Z1377, 共含有18个代换片段, 平均代换长度2.95 Mb, 携带多个育种有利的性状。Z1377的株高增加是节间数增多2节及各节间和穗长共同增加的结果。Z1377的株高由一个主效QTL (qPH3)和一个微效QTL控制, 且主要由qPH3负责, 呈显性单基因遗传, 为首次报道。Z1377共携带16个重要性状的QTL, 其中8个已被克隆。该研究对qPH3的精细定位和克隆以及为进一步选育含2~3个代换片段的中高优良染色体片段代换系并应用于育种奠定了基础。

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] Wang Y H, Li J Y . The plant architecture of rice (Oryza sativa L.)
Plant Mol Biol, 2005,59:75-84
[本文引用: 1]

[2] Monna L, Kitazawa N, Yoshino R, Suzuki J, Masuda H, Maehara Y, Tanji M, Sato M, Nasu S, Minobe Y . Positional cloning of rice semidwarfing gene, sd-1: rice “green revolution gene” encodes a mutant enzyme involved in gibberellin synthesis
DNA Res, 2002,9:11-17
[本文引用: 1]

[3] Tong H N, Xiao Y H, Liu D P, Gao S P, Liu L C, Yin Y H, Jin Y, Qian Q, Chu C C . Brassinosteroid regulates cell elongation by modulating gibberellin metabolism in rice
Plant Cell, 2014,26:4376-4393
DOI:10.1105/tpc.114.132092URLPMID:25371548 [本文引用: 2]
Brassinosteroid (BR) and gibberellin (GA) are two predominant hormones regulating plant cell elongation. A defect in either of these leads to reduced plant growth and dwarfism. However, their relationship remains unknown in rice (Oryza sativa). Here, we demonstrated that BR regulates cell elongation by modulating GA metabolism in rice. Under physiological conditions, BR promotes GA accumulation by regulating the expression of GA metabolic genes to stimulate cell elongation. BR greatly induces the expression of D18/GA3ox-2, one of the GA biosynthetic genes, leading to increased GA1 levels, the bioactive GA in rice seedlings. Consequently, both d18 and loss-of-function GA-signaling mutants have decreased BR sensitivity. When excessive active BR is applied, the hormone mostly induces GA inactivation through upregulation of the GA inactivation gene GA2ox-3 and also represses BR biosynthesis, resulting in decreased hormone levels and growth inhibition. As a feedback mechanism, GA extensively inhibits BR biosynthesis and the BR response. GA treatment decreases the enlarged leaf angles in plants with enhanced BR biosynthesis or signaling. Our results revealed a previously unknown mechanism underlying BR and GA crosstalk depending on tissues and hormone levels, which greatly advances our understanding of hormone actions in crop plants and appears much different from that in Arabidopsis thaliana.

[4] Wu J H, Zhu C F, Pang J H, Zhang X R, Yang C L, Xia G X, Tian Y C, He C Z . OsLOL1, a C2C2-type zinc finger protein, interacts with OsbZIP58 to promote seed germination through the modulation of gibberellin biosynthesis in Oryza sativa
Plant J, 2014,80:1118-1130
[本文引用: 1]

[5] Hong Z, Ueguchi-Tanaka M, Shimizu-Sato S, Inukai Y, Fujioka S, Shimada Y, Takatsuto S, Agetsuma M, Yoshida S, Watanabe Y, Uozu S, Kitano H, Ashikari M, Matsuoka M . Loss-of-function of a rice brassinosteroid biosynthetic enzyme, C-6 oxidase, prevents the organized arrangement and polar elongation of cells in the leaves and stem
Plant J, 2002,32:495-508
DOI:10.1046/j.1365-313X.2002.01438.xURL [本文引用: 2]

[6] Tanabe S, Ashikari M, Fujioka S, Takatsuto S, Yoshida S, Yano M, Yoshimura A, Kitano H, Matsuoka M, Fujisawa Y, Kato H, Iwasaki Y . A novel cytochrome P450 is implicated in brassinosteroid biosynthesis via the characterization of a rice dwarf nutant, dwarf11, with reduced seed length
Plant Cell, 2005,17:776-790
[本文引用: 2]

[7] Tong H N, Jin Y, Liu W B, Li F, Fang J, Yin Y H, Qian Q, Zhu L H, Chu C C . DWARF AND LOW-TILLERING, a new member of the GRAS family, plays positive roles inbrassinosteroid singaling in rice
Plant J, 2009,58:803-816
[本文引用: 1]

[8] Ito S, Kitahata N, Umehara M, Hanada A, Kato A, Ueno K, Mashiguchi K, Kyozuka J, Yoneyama K, Yamaguchi S, Asami T . A new lead chemical for strigolactone biosynthesis inhibitors
Plant Cell Physiol, 2010,51:1143-1150
DOI:10.1093/pcp/pcq077URLPMID:20522488 [本文引用: 1]
Several triazole-containing chemicals have previously been shown to act as efficient inhibitors of cytochrome P450 monooxygenases. To discover a strigolactone biosynthesis inhibitor, we screened a chemical library of triazole derivatives to find chemicals that induce tiller bud outgrowth of rice seedlings. We discovered a triazole-type chemical, TIS13 [2,2-dimethyl-7-phenoxy-4-(1H-1,2,4-triazol-1-yl)heptan-3-ol], which induced outgrowth of second tiller buds of wild-type seedlings, as observed for non-treated strigolactone-deficientd10mutant seedlings. TIS13 treatment reduced strigolactone levels in both roots and root exudates in a concentration-dependent manner. Co-application of GR24, a synthetic strigolactone, with TIS13 canceled the TIS13-induced tiller bud outgrowth. Taken together, these results indicate that TIS13 inhibits strigolactone biosynthesis in rice seedlings. We propose that TIS13 is a new lead compound for the development of specific strigolactone biosynthesis inhibitors.

[9] 赵芳明, 郭超, 魏霞, 杨正林, 凌英华, 桑贤春, 王楠, 张长伟, 李云峰, 何光华 . 日本晴与5个优良恢复系的多态性标记筛选及遗传差异分析
西南大学学报(自然科学版), 2016,38(11):1-7
[本文引用: 1]

Zhao F M, Guo C, Wei X, Yang Z L, Ling Y H, Sang X C, Wang N, Zhang C W, Li Y F, He G H . Polymorphic SSR markers screening and genetic difference analysis between Nipponbare and five excellent restorer line
J Southwest Univ (Nat Sci Edn), 2016,38(11):1-7 (in Chinese with English abstract)
[本文引用: 1]

[10] 向佳, 李燕, 樊亚伟, 许军红, 郑丽媛, 何光华, 杨正林, 王楠, 赵芳明 . 一个具有主效晚抽穗基因的水稻染色体片段代换系的鉴定、形态分析及Ehd4-2定位
作物学报, 2015,41:683-691
[本文引用: 3]

Xiang J, Li Y, Fan Y W, Xu J H, Zheng L Y, He G H, Yang Z L, Wang N, Zhao F M . Identification and morphological analysis of a rice chromosome segment substitution line carrying a major effect gene for late heading date and mapping of Ehd4-2
Acta Agron Sin, 2015,41:683-691 (in Chinese with English abstract)
[本文引用: 3]

[11] Paterson A H, Damon S, Hewitt J D, Zamir D, Rabinowitch H D, Lincoln S E, Lander E S, Tanksley S D . Mendelian factors underlying quantitative traits in tomato: comparison across species, generations, and environments
Genetics, 1991,127:181-197
[本文引用: 1]

[12] Zhao F M, Tan Y, Zheng L Y, Zhou K, He G H, Ling Y H, Zhang L H, Xu S Z . Identification of rice chromosome segment substitution line Z322-1-10 and mapping QTL for agronomic traits from the F₃ population
Cereal Res Commun, 2016,44:370-380
DOI:10.1556/0806.44.2016.022URL [本文引用: 1]

[13] Furuta T, Uehara K , Angeles-Shim R B, Shim J, Ashikari M, Takashi T. Development and evaluation of chromosome segment substitution lines (CSSLs) carrying chromosome segments derived from Oryza rufipogon in the genetic background of Oryza sativa L
Breed Sci, 2014,63:468-475
[本文引用: 1]

[14] 王坚, 赵开军, 乔枫, 杨生龙 . OsGA20ox2不同长度RNAi片段对水稻株高等农艺性状的遗传效应
作物学报, 2012,38:632-638
[本文引用: 1]

Wang J, Zhao K J, Qiao F, Yang S L . Genetic effects of different RNA interference fragments from OsGA20ox2 on plant height and other agronomic traits in rice
Acta Agron Sin, 2012,38:632-638 (in Chinese with English abstract)
[本文引用: 1]

[15] Zhu Y Y, Nomura T, Xu Y H, Zhang Y Y, Peng Y, Mao B Z, Hanada A, Zhou H C, Wang R X, Li P J, Zhu X D, Mander L N, Kamiya Y, Yamaguchi S, He Z H . ELONGATED UPPERMOST INTERNODE encodes a cytochrome P450 monooxygenase that epoxidizes gibberellins in a novel deactivation reaction in rice
Plant Cell, 2006,18:442-456
[本文引用: 1]

[16] Zhang Y, Lan H X, Shao Q L, Wang R Q, Chen H, Tang H J, Zhang H S, Huang J . An A20/AN1-type zinc finger protein modulates gibberellins and abscisic acid contents and increases sensitivity to abiotic stress in rice (Oryza sativa)
J Exp Bot, 2016,67:315-326
[本文引用: 1]

[17] Chen J, Gao H, Zheng X M, Jin M N, Weng J F, Ma J, Ren Y L, Zhou K N, Wang Q, Wang J, Wang J L, Zhang X, Cheng Z J, Wu C Y, Wang H Y, Wan J M . An evolutionarily conserved gene, FUWA, plays a role in determining panicle architecture, grain shape and grain weight in rice
Plant J, 2015,83:427-438
[本文引用: 1]

[18] Li M, Tang D, Wang K J, Wu X R, Lu L L, Yu H X, Gu M H, Yan C J, Cheng Z K . Mutations in the F-box gene LARGER PANICLE improve the panicle architecture and enhance the grain yield in rice
Plant Biotechnol J, 2011,9:1002-1013
[本文引用: 1]

[19] Song X J, Huang W, Shi M, Zhu M Z, Lin H X . A QTL for rice grain width and weight encodes a previously unknown RING-type E3 ubiquitin ligase
Nat Genet, 2007,39:623-630
DOI:10.1038/ng2014URL [本文引用: 1]

[20] Heang D, Sassa H . Antagonistic actions of HLH/bHLH proteins are involved in grain length and weight in rice
PLoS One, 2012,7(2):e31325
DOI:10.1371/journal.pone.0031325URLPMID:3283642 [本文引用: 1]
Grain size is a major yield component in rice, and partly controlled by the sizes of the lemma and palea. Molecular mechanisms controlling the sizes of these organs largely remain unknown. In this study, we show that an antagonistic pair of basic helix-loop-helix (bHLH) proteins is involved in determining rice grain length by controlling cell length in the lemma/palea. Overexpression of an atypical bHLH, named POSITIVE REGULATOR OF GRAIN LENGTH 1 (PGL1), in lemma/palea increased grain length and weight in transgenic rice. PGL1 is an atypical non-DNA-binding bHLH and assumed to function as an inhibitor of a typical DNA-binding bHLH through heterodimerization. We identified the interaction partner of PGL1 and named it ANTAGONIST OF PGL1 (APG). PGL1 and APG interacted in vivo and localized in the nucleus. As expected, silencing of APG produced the same phenotype as overexpression of PGL1, suggesting antagonistic roles for the two genes. Transcription of two known grain-length-related genes, GS3 and SRS3, was largely unaffected in the PGL1-overexpressing and APG-silenced plants. Observation of the inner epidermal cells of lemma revealed that are caused by increased cell length. PGL1-APG represents a new grain length and weight-controlling pathway in which APG is a negative regulator whose function is inhibited by PGL1.

[21] Taguchi-Shiobara F, Kawagoe Y, Kato H, Onodera H, Tagiri A, Hara N, Miyao A, Hirochika H, Kitano H, Yano M, Toki S . A loss-of-function mutation of rice DENSE PANICLE 1 causes semi-dwarfness and slightly increased number of spikelets
Breed Sci, 2011,61:17-25
[本文引用: 1]

[22] Huang X Z, Qian Q, Liu Z B, Sun H Y, He S Y, Luo D, Xia G M, Chu C C, Li J Y, Fu X D . Natural variation at the DEP1 locus enhances grain yield in rice
Nat Genet, 2009,41:494-497
[本文引用: 1]

[23] 刘胜男, 张华, 柳絮, 李广贤, 杨永义, 姚方印 . 水稻株高和产量相关性状的QTL定位
山东农业科学, 2015,47(4):8-12
URL [本文引用: 1]

Liu S N, Zhang H, Liu X, Li G X, Yang Y Y, Yao F Y . QTL mapping of plant height and yield-related traits of rice
Shandong Agric Sci, 2015,47(4):8-12
URL [本文引用: 1]

[24] Xue W, Xing Y, Weng X, Zhao Y, Tang W, Wang L, Zhou H, Yu S, Xu C, Li X, Zhang Q . Natural variation in Ghd7 is an important regulator of heading date and yield potential in rice
Nat Genet, 2008,40:761-767
DOI:10.1038/ng.143URL [本文引用: 1]

[25] Jones D F . Dominance of linked factors as a means of accounting for heterosis
Genetics, 1997,2:466-479
[本文引用: 1]

[26] Cai W, Morishima H . QTL clusters reflect character associations in wild and cultivated rice
Theor Appl Genet, 2002,104:1217-1228
DOI:10.1007/s00122-001-0819-7URLPMID:12582574 [本文引用: 1]
The genetic basis of character association related to differentiation found in the primary gene pool of rice was investigated based on the genomic distribution of quantitative trait loci (QTLs). Major evolutionary trends in cultivated rice of Asiatic origin ( Oryza sativa ) and its wild progenitor ( O. rufipogon ) are: (1) differentiation from wild to domesticated types (domestication), (2) ecotype differentiation between the perennial and annual types in wild races, and (3) the Indica versus Japonica type differentiation in cultivated races. Using 125 recombinant inbred lines (RILs) derived from a cross between an Indica cultivar of O. sativa and a strain of O. rufipogon carrying some Japonica-like characteristics, we mapped 147 markers, mostly RFLPs, on 12 chromosomes. Thirty-seven morphological and physiological quantitative traits were evaluated, and QTLs for 24 traits were detected. The mapped loci showed a tendency to form clusters that are composed of QTLs of the domestication-related traits as well as Indica/Japonica diagnostic traits. QTLs for perennial/annual type differences did not cluster. This cluster phenomenon could be considered "multifactorial linkages" followed by natural selection favoring co-adapted traits. Further, it is possible that the clustering phenomenon is partly due to pleiotropy of some unknown key factor(s) controlling various traits through diverse metabolic pathways. Chromosomal regions where QTL clusters were found coincided with the regions harboring genes or gene blocks where the frequency of cultivar-derived alleles in RILs is higher than expected. This distortion may be partly due to unconscious selection favoring cultivated plant type during the establishment of RILs.