以三明显性核不育系转导稻瘟病抗性基因<i>Pi9</i>改良恢复系的效果

邓则勤; 黄显波; 林成豹; 唐江霞; 叶仰东; 苏荣理; 梁水金

doi:10.19303/j.issn.1008-0384.2020.01.002

以三明显性核不育系转导稻瘟病抗性基因Pi9改良恢复系的效果

doi: 10.19303/j.issn.1008-0384.2020.01.002

福建省三明市农业科学研究院，福建　三明　365051

基金项目: 福建省科技计划农业引导性项目（2017N0050）；福建省种业创新与产业化工程项目（fjzycxny2017001）

详细信息

作者简介:
邓则勤（1972−），男，副研究员，研究方向：水稻育种（E-mail: dengzeqin1998@163.com）

通讯作者:
黄显波（1964−），男，研究员，研究方向：水稻育种（E-mail: huangxb_2000@163.com）

中图分类号: S 511
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出版历程
- 收稿日期: 2019-11-11
- 修回日期: 2020-01-02
- 刊出日期: 2020-01-01

Improvement of Disease-resistance of Restorer Rice Lines by Transducing Genes Pi9 onto Sanming Dominant Male Sterile Lines

Sanming Academy of Agricultural Sciences, Sanming, Fujian　365051, China

摘要

摘要: 目的将广谱抗稻瘟病基因Pi9导入恢复系双抗明占，以提高其组合抗病能力。方法通过构建Pi9基因的三明显性核不育系载体作中间“架桥”，结合分子标记辅助选择，每个回交世代选择有Pi9标记基因带型的显性核不育株与双抗明占回交，至BC₄F₁代筛选获得检测有Pi9标记基因带型可育株自交纯合Pi9基因，获得卸载了核不育基因的双抗明占抗病近等基因系。结果获选的6个改良系经田间抗性鉴定有5个抗稻瘟病能力得到显著提升，进行配合力测验及农艺性状比较分析，改良系Y25-1一般配合力表现最好，抗病能力强，主要农艺性状与双抗明占相近，可作为抗病的双抗明占推广应用。结论通过水稻三明显性核不育系作中间“架桥”，导入Pi9基因可有效提高双抗明占及其组合抗稻瘟病能力。
- 水稻 /
- 三明显性核不育系 /
- 载体 /
- 转导 /
- Pi9 /
- 双抗明占
Abstract: Objective To transfer the broad-spectrum rice blast resistance gene, Pi9, to the restorer line Shuangkangmingzhan for improving its disease resistance. Method The Sanming dominant male sterile line, a carrier of the Pi9 gene, was used as the intermediate “bridge” for the gene transduction experiment. Using the molecular marker assistance selection method, from each backcross generation the male sterile plants with Pi9 marker genotype were selected to backcross- mate with Shuangkangmingzhan until the self-homozygous Pi9 gene marker in the fertile plant was detected. Then, from the selected plant in the BC₄F₁ generation of the Shuangkangmingzhan disease-resistant isogenic line, the target sterile gene was obtained. Result Of the 6 improved lines selected, 5 were identified by the field resistance verification with significantly improved resistance to rice blast. The combining abilities and agronomic traits of the lines were compared and analyzed to show that the improved line Y25-1 possessed the best general combining ability with a strong blast resistance. Since its main agronomic traits were similar to those of Shuangkangmingzhan, it could conceivably be considered a surrogate of the disease-resistant Shuangkangmingzhan for breeding purpose. Conclusion For an improved blast resistance on rice, the successful transferring Pi9 gene from Shuangkangmingzhan onto Y25-1 through the use of the Sanming dominant male sterile line as an intermediate “bridge” provided a new venue for the breeding.
- rice /
- Sanming dominant male sterility /
- carrier /
- transduction /
- Pi9 /
- Shuangkangmingzhan

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表 1 三明显性核不育基因与Pi9基因不同回交世代分离比例

Table 1. Separation rate of Sanming dominant male sterile gene and Pi9 in backcross generations

回交世代 Backcross generation	Pi9不育株数 Pi9 sterile plants	pi9不育株数 pi9 sterile plants	Pi9可育株数 Pi9 fertile plants	pi9可育株数 pi9 fertile plants	期望分离比 Expectation separation ratio	X ²	P_0.05
BC₁F₁	94	111	105	91	1∶1∶1∶1	2.621	7.815
BC₂F₁	102	108	99	114	1∶1∶1∶1	1.255	7.815
BC₃F₁	34	45	37	44	1∶1∶1∶1	2.159	7.815
注：X ²_0.05，3=7.815

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表 2 双抗明占与改良株系的稻瘟病抗性表现

Table 2. Blast resistances of Shuangkangmingzhan and selected plants

亲本 Parent	苗瘟 Seeding Blast					叶稻瘟 Leaf Blast					穗颈瘟 Panicle Blast					抗性综评 Comprehensive evaluation
	Ⅰ		Ⅱ		最高级别 Level	Ⅰ		Ⅱ		最高级别 Level	Ⅰ		Ⅱ		最高级别 Level
	发病率 Prevalence/%	级别 Level	发病率 Prevalence/%	级别 Level	最高级别 Level	发病率 Prevalence/%	级别 Level	发病率 Prevalence/%	级别 Level	最高级别 Level	发病率 Prevalence/%	级别 Level	发病率 Prevalence/%	级别 Level	最高级别 Level
双抗明占 Shuangkang mingzhan	8	4	7	4	4	7	4	5	4	4	11	5	12	5	5	MS
Y14-1	5	4	8	4	4	2	3	3	4	4	7	3	8	3	3	MR
Y17-1	7	4	6	4	4	3	4	5	4	4	17	5	14	5	5	MS
Y18-1	4	3	2	3	3	0	0	0	0	0	3	1	4	1	1	R
Y22-1	17	5	24	6	6	4	3	3	4	4	3	1	2	1	1	MR
Y25-1	6	4	5	4	4	0	0	0	0	0	4	1	5	1	1	R
Y28-1	3	3	2	3	3	0	0	0	0	0	2	1	3	1	1	R
感病对照 Control	75	9	78	9	9	56	7	63	7	7	76	9	85	9	9	HS
注：MS：中感，HS：高感，MR：中抗，R：抗。 Note: MS-medium sensitive, HS-highly sensitive, MR-medium resistant, R-resistant.

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表 3 亲本农艺性状的一般配合力的相对效应

Table 3. Relative effect of general combining ability on agronomic traits of 11 parents

亲本 Parent	株高 Plant height	抽穗期 Heading date	单株有效穗 Effective panicle	每穗实粒数 Grains per panicle	每穗总粒数 Spikelets per panicle	结实率 Seed setting rate	千粒重 1 000-grain weight	单株产量 Grain yield per plant
赣香 A Ganxiang A	6.56	2.75	14.22	6.44	1.84	4.54	11.25	33.66
泸香618A Luxiang 618A	−4.19	−3.65	−4.35	−9.09	−3.05	−6.25	−1.51	−15.35
T108S	1.55	0.34	−6.54	−1.00	1.83	−2.76	−6.14	−13.76
明1A Ming 1A	−3.92	0.56	−3.33	3.64	−0.61	4.47	−3.60	−4.55
双抗明占 Shuangkang mingzhan	−0.09	0.73	9.03	0.96	0.00	1.59	−1.64	9.46
Y14-1	0.00	0.04	−14.77	16.54	14.90	0.56	−0.72	1.86
Y17-1	−0.55	−0.55	0.84	−4.01	−4.25	0.56	0.36	−2.19
Y18-1	1.07	−0.74	−2.49	−2.82	−3.77	1.02	1.93	−4.80
Y22-1	0.26	0.24	−5.81	−0.61	−0.67	0.33	−1.08	−6.74
Y25-1	0.08	0.24	10.82	−7.38	−3.66	−3.70	0.80	2.57
Y28-1	−0.77	0.04	2.38	−2.67	−2.56	−0.36	0.35	−0.16

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表 4 改良系及双抗明占主要农艺性状表现

Table 4. Agronomic traits of improved lines and Shuangkangmingzhan

亲本 Parent	双抗明占 Shuangkang mingzhan	Y14-1	Y17-1	Y18-1	Y22-1	Y25-1	Y28-1
株高 Plant height/cm	100.3±1.2 c	104.6±0.8 a	100.0±2.1 c	101.3±0.6 bc	100.5±1.9 c	104.9±0.3 a	103.3±0.5 ab
剑叶长 Flag leaf length/cm	24.0±4.4 a	19.7±2.3 a	19.7±3.4 a	20.7±3.5 a	21.0±1.5 a	19.3±2.2 a	21.4±0.5 a
剑叶宽 Flag leaf width/cm	1.7±0.1 a	1.7±0.1 a	1.6±0.1 a	1.7±0.1 a	1.6±0.1 a	1.6±0.1 a	1.7±0.2 a
倒二叶长 Inverted second leaf length/cm	35.8±4.7 a	32.5±4.1 a	31.5±4.9 a	30.6±1.6 a	31.8±3.5 a	32.5±3.5 a	33.4±1.0 a
倒二叶宽 Inverted second leaf width/cm	1.4±0.1 a	1.3±0.1 ab	1.2±0.1 b	1.2±0.1 b	1.2±0.1 b	1.2±0.1 b	1.3±0.1 ab
穗长 Panicel length/cm	24.9±0.9 a	24.8±1.3 a	25.7±0.9 a	24.6±0.7 a	24.5±1.0 a	25.6±0.9 a	25.0±1.4 a
第1节间长 Length of the first internode/cm	1.7±0.3 b	3.0±0.3 a	3.2±1.0 a	2.8±0.4 ab	2.0±0.4 ab	2.5±0.3 ab	2.6±0.4 ab
第2节间长 Length of the second internode/cm	6.9±1.5 b	11.0±1.0 a	9.2±0.6 ab	8.3±0.9 b	8.6±2.1 ab	8.1±1.5 b	8.2±1.7 b
第3节间长 Length of the third internode/cm	10.2±0.2 b	12.7±0.1 a	12.7±0.4 a	12.9±0.6 a	12.4±1.2 a	12.8±0.3 a	12.6±0.6 a
第4节间长 Length of the fourth internode/cm	18.5±1.0 a	19.2±1.4 a	18.7±0.5 a	19.0±0.4 a	19.4±0.6 a	19.2±1.2 a	19.4±0.8 a
第5节间长 Length of the fifth internode/cm	36.9±3.2 a	34.5±0.8 ab	32.9 ±3.0 b	34.4±0.8 ab	34.2±1.1 ab	34.6±0.5 ab	34.6±0.6 ab
茎粗 Stem diameter/cm	0.57±0.1 a	0.55±0.1 a	0.57±0.1 a	0.53±0.1 a	0.53±0.1 a	0.57±0.1 a	0.53±0.1 a
单株有效穗 Effective panicle/穗	11.1±0.3 a	8.9±0.7 cd	9.0±0.4 cd	9.4±0.3 bc	8.9±0.3 cd	10.2±0.3 ab	8.2±0.7 d
每穗总粒数 Spikelets per panicle/粒	156.7±3.2 cd	163.1±1.6 b	168.5±2.3 a	158.6±0.7 c	151.7±0.9 e	153.0±1.3 de	172.4±2.4 a
每穗实粒数 Grains per panicle/粒	130.6±0.9 a	128.3±1.1 a	130.7±1.7 a	128.3±1.0 a	116.6±1.8 b	128.4±1.5 a	127.8±0.7 a
结实率 Seed setting rate/%	83.5±1.7 a	79.5±1.8 bc	77.5±1.1 c	80.5±0.9 b	77.4±1.0 c	84.6±0.8 a	74.4±0.6 d
千粒重1 000-grain weight/g	24.73±0.1 a	24.50±0.2 ab	23.94±0.1 cd	24.21±0.1 bc	23.70±0.1 d	24.36±0.2 b	23.91±0.1 cd
注：同行数据后不同小写字母表示差异显著（P＜0.05）。 Note: Data with different lowercase letters on a same line indicate significant difference at P＜0.05.

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