浙江近年育成粳稻新品种(系)部分抗病虫基因的分子检测与育种应用

doi:10.3969/j.issn.1004-1524.2021.07.01

浙江农业学报 ›› 2021, Vol. 33 ›› Issue (7): 1159-1169.DOI: 10.3969/j.issn.1004-1524.2021.07.01

浙江近年育成粳稻新品种(系)部分抗病虫基因的分子检测与育种应用

黄宣(), 金林灿, 叶朝辉, 姜洁锋, 施贤波

宁波市农业科学研究院作物研究所,浙江宁波 315040

收稿日期:2020-12-29 出版日期:2021-07-25 发布日期:2021-08-06
作者简介:黄宣(1981—),男,河南信阳人,硕士,农艺师,主要从事常规晚粳稻新品种选育与推广研究。E-mail: huangx0913@126.com
基金资助:
宁波市科技局中央引导地方科技发展专项(2019C10106);浙江省农业新品种选育重大科技专项(2016C02050-5)

Molecular detection and breeding application of some disease and insect resistance genes of japonica rice varieties/lines recently developed in Zhejiang Province

HUANG Xuan(), JIN Lincan, YE Chaohui, JIANG Jiefeng, SHI Xianbo

Institute of Crops, Ningbo Academy of Agricultural Sciences, Ningbo 315040,China

Received:2020-12-29 Online:2021-07-25 Published:2021-08-06

摘要/Abstract

摘要：

为明确浙江省近年育成常规晚粳稻品种(系)的稻瘟病、白叶枯病、褐飞虱等抗性基因的分布情况,利用分子标记检测分析了11个抗稻瘟病基因(Pita、Pib、Pigm、Pikh/Pi54、Pi9、Pi1、Pi2、Pi5、Pit、Pb1、Pikm),2个抗白叶枯基因(Xa21、Xa23)和2个抗褐飞虱基因(Bph14、Bph15)在61个晚粳稻品种(系)分布情况。结果表明:稻瘟病抗性基因检出率最高的为Pib,分布频率为91.80%,Pikm、Pi2、Pita的分布频率分别为68.85%、67.21%、63.93%,Pit、Pi5、Pi9、Pikh/Pi54检出频率分别为34.43%、32.79%、26.23%、24.59%;Pb1、pigm检出率较低,分别为6.56%、1.64%;Pi1基因未检出;抗褐飞虱基因Bph14、Bph15检出率分别为6.56%、11.48%;抗白叶枯基因Xa21、Xa23在61个被检测品种(系)中无分布。利用分子标记辅助选择将Xa23、Bph14、Bph15导入粳稻中并验证了抗性,结果表明,以上基因均具有较好的抗性。初步明确了浙江省新育成61个常规晚粳稻品种(系)的抗病虫基因分布情况,研究结果将为今后利用分子标记辅助技术将目标抗性基因转移到待改良品种中培育广谱多抗水稻新品种提供依据。

关键词: 粳稻, 稻瘟病, 白叶枯病, 褐飞虱, 分子标记, 抗性基因检测

Abstract:

In order to clarify the distribution of some disease and insect resistance genes of japonica rice varieties/lines recently developed in Zhejiang Province, in this study, the distribution of blast resistance gene Pita, Pib, Pigm,Pi9, Pi1, Pikh/Pi54, Pi2, Pi5, Pit, Pb1, Pikm and BPH resistance gene Bph14, Bph15, bacterial blight resistance gene Xa21, Xa23 in 61 japonica rice varieties/lines were analyzed.The results showed that the highest detection frequency of rice blast resistance gene was Pib, the distribution frequency was 91.80%, the distribution frequency of Pikm, Pi2 and Pita were 68.85%, 67.21%, 63.93%, respectively; the distribution frequency of Pit, Pi5, Pi9, Pikh/Pi54 were 34.43%, 32.79%, 26.23% and 24.59%, respectively. The distribution frequency of Pb1 and Pigm were 6.56%, 1.64%,respectively. The Pi1 was not distributed in 61 tested varieties/lines. The distribution frequency of Bph14 and Bph15 were 6.56% and 11.48%, respectively. The bacterial blight resistance gene Xa21 and Xa23were not distributed in 61 tested varieties/lines. In this study, Xa23, Bph14 and Bph15 were introduced into japonica rice by marker assisted selection, and the results showed that the above genes had good resistance. In this study, the distribution of resistance genes of 61 japonica rice varieties/lines in Zhejiang Province were preliminarily established, the results of this study will provide a scientific basis for transferring the target resistance genes into the varieties to be improved and breeding new varieties with broad-spectrum and multi resistance.

Key words: japonica rice, blast, bacterial blight, rice brown planthopper, molecular marker, resistance gene detection

中图分类号:

黄宣, 金林灿, 叶朝辉, 姜洁锋, 施贤波. 浙江近年育成粳稻新品种(系)部分抗病虫基因的分子检测与育种应用[J]. 浙江农业学报, 2021, 33(7): 1159-1169.

HUANG Xuan, JIN Lincan, YE Chaohui, JIANG Jiefeng, SHI Xianbo. Molecular detection and breeding application of some disease and insect resistance genes of japonica rice varieties/lines recently developed in Zhejiang Province[J]. Acta Agriculturae Zhejiangensis, 2021, 33(7): 1159-1169.

图/表 16

参考文献 35

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[29]	陈涛, 孙旭超, 张善磊, 等. 稻瘟病广谱抗性基因Pigm特异性分子标记的开发和应用[J]. 中国水稻科学, 2020, 34(1):28-36.
	CHEN T, SUN X C, ZHANG S L, et al. Development and verification of specific molecular markers for Pigm gene associated with broad-spectrum resistance to rice blast[J]. Chinese Journal of Rice Science, 2020, 34(1):28-36.(in Chinese with English abstract)
[30]	于苗苗, 戴正元, 潘存红, 等. 广谱稻瘟病抗性基因Pigm和Pi2的抗谱差异及与Pi1的互作效应[J]. 作物学报, 2013, 39(11):1927-1934.
	YU M M, DAI Z Y, PAN C H, et al. Resistance spectrum difference between two broad-spectrum blast resistance genes, Pigm and Pi2, and their interaction effect on Pi1[J]. Acta Agronomica Sinica, 2013, 39(11):1927-1934.(in Chinese with English abstract) DOI URL
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选育单位Breeding institute	品种(系)Varieties/lines
中国水稻研究所 China National Rice Research Institute	春江157 Chunjiang157^、中嘉8号Zhongjia8hao^、春江糯8号Chunjiangnuo8、春江糯7号 Chunjiangnuo7、春江195 Chunjiang195、春江191 Chunjiang191、春江190 Chunjiang190、春江188 Chunjiang188、春江183 Chunjiang183、春江171 Chunjiang171
浙江省农业科学院 Zhejiang Academy of Agricultural Sciences	浙糯106 Zhenuo106^、浙粳96 Zhejing96^、ZH19-6、ZH19-12、ZH19-02、ZH18-47、ZH18-43、ZH18-117、ZH17-122、ZH16-98、R9079、R202、R201、R192、R1825、R182、R1751
宁波市农业科学研究院 Ningbo Academy of Agricultural Sciences	宁84 Ning84^,宁88 Ning88^、宁81 Ning81^、宁82 Ning82^、宁39 Ning39,甬粳19-46 Yongjing19-46、甬粳18-60 Yongjing18-60
嘉兴市农业科学研究院 Jiaxing Academy of Agricultural Sciences	秀水14 Xiushui14^、秀水134 Xiushui134^、秀水121 Xiushui121^、嘉67 Jia67^、天微香粳 Tianweijing、嘉禾628 Jiahe218、嘉60 Jia60、嘉16-48 Jia16-48、嘉124 Jia124、丙19-339 Bing19-339、丙19-26 Bing19-26、丙18-979 Bing18-979、丙18-927 Bing18-927、丙17-204 Bing17-204、丙17-02 Bing17-02、丙16-135 Bing16-135、丙16-111 Bing16-111、丙10-544 Bing10-544
湖州市农业科学研究院 Huzhou Academy of Agricultural Sciences	HZ16-78、HZ19-1137B、HZ19-1137C
绍兴市农业科学研究院 Shaoxing Academy of Agricultural Sciences	绍糯9714 Shaonuo9714^、绍粳18 Shaojing18^、绍粳19-6 Shaojing19-6 、绍粳19-34 Shaojing19-34、绍粳19-19 Shaojing19-19
台州市农业科学研究院 Taizhou Academy of AgriculturalSciences	台19-84 Tai19-84

选育单位Breeding institute	品种(系)Varieties/lines
中国水稻研究所 China National Rice Research Institute	春江157 Chunjiang157^、中嘉8号Zhongjia8hao^、春江糯8号Chunjiangnuo8、春江糯7号 Chunjiangnuo7、春江195 Chunjiang195、春江191 Chunjiang191、春江190 Chunjiang190、春江188 Chunjiang188、春江183 Chunjiang183、春江171 Chunjiang171
浙江省农业科学院 Zhejiang Academy of Agricultural Sciences	浙糯106 Zhenuo106^、浙粳96 Zhejing96^、ZH19-6、ZH19-12、ZH19-02、ZH18-47、ZH18-43、ZH18-117、ZH17-122、ZH16-98、R9079、R202、R201、R192、R1825、R182、R1751
宁波市农业科学研究院 Ningbo Academy of Agricultural Sciences	宁84 Ning84^,宁88 Ning88^、宁81 Ning81^、宁82 Ning82^、宁39 Ning39,甬粳19-46 Yongjing19-46、甬粳18-60 Yongjing18-60
嘉兴市农业科学研究院 Jiaxing Academy of Agricultural Sciences	秀水14 Xiushui14^、秀水134 Xiushui134^、秀水121 Xiushui121^、嘉67 Jia67^、天微香粳 Tianweijing、嘉禾628 Jiahe218、嘉60 Jia60、嘉16-48 Jia16-48、嘉124 Jia124、丙19-339 Bing19-339、丙19-26 Bing19-26、丙18-979 Bing18-979、丙18-927 Bing18-927、丙17-204 Bing17-204、丙17-02 Bing17-02、丙16-135 Bing16-135、丙16-111 Bing16-111、丙10-544 Bing10-544
湖州市农业科学研究院 Huzhou Academy of Agricultural Sciences	HZ16-78、HZ19-1137B、HZ19-1137C
绍兴市农业科学研究院 Shaoxing Academy of Agricultural Sciences	绍糯9714 Shaonuo9714^、绍粳18 Shaojing18^、绍粳19-6 Shaojing19-6 、绍粳19-34 Shaojing19-34、绍粳19-19 Shaojing19-19
台州市农业科学研究院 Taizhou Academy of AgriculturalSciences	台19-84 Tai19-84

选育单位Breeding institute	品种(系)Varieties/lines
中国水稻研究所 China National Rice Research Institute	春江157 Chunjiang157^、中嘉8号Zhongjia8hao^、春江糯8号Chunjiangnuo8、春江糯7号 Chunjiangnuo7、春江195 Chunjiang195、春江191 Chunjiang191、春江190 Chunjiang190、春江188 Chunjiang188、春江183 Chunjiang183、春江171 Chunjiang171
浙江省农业科学院 Zhejiang Academy of Agricultural Sciences	浙糯106 Zhenuo106^、浙粳96 Zhejing96^、ZH19-6、ZH19-12、ZH19-02、ZH18-47、ZH18-43、ZH18-117、ZH17-122、ZH16-98、R9079、R202、R201、R192、R1825、R182、R1751
宁波市农业科学研究院 Ningbo Academy of Agricultural Sciences	宁84 Ning84^,宁88 Ning88^、宁81 Ning81^、宁82 Ning82^、宁39 Ning39,甬粳19-46 Yongjing19-46、甬粳18-60 Yongjing18-60
嘉兴市农业科学研究院 Jiaxing Academy of Agricultural Sciences	秀水14 Xiushui14^、秀水134 Xiushui134^、秀水121 Xiushui121^、嘉67 Jia67^、天微香粳 Tianweijing、嘉禾628 Jiahe218、嘉60 Jia60、嘉16-48 Jia16-48、嘉124 Jia124、丙19-339 Bing19-339、丙19-26 Bing19-26、丙18-979 Bing18-979、丙18-927 Bing18-927、丙17-204 Bing17-204、丙17-02 Bing17-02、丙16-135 Bing16-135、丙16-111 Bing16-111、丙10-544 Bing10-544
湖州市农业科学研究院 Huzhou Academy of Agricultural Sciences	HZ16-78、HZ19-1137B、HZ19-1137C
绍兴市农业科学研究院 Shaoxing Academy of Agricultural Sciences	绍糯9714 Shaonuo9714^、绍粳18 Shaojing18^、绍粳19-6 Shaojing19-6 、绍粳19-34 Shaojing19-34、绍粳19-19 Shaojing19-19
台州市农业科学研究院 Taizhou Academy of AgriculturalSciences	台19-84 Tai19-84

选育单位Breeding institute	品种(系)Varieties/lines
中国水稻研究所 China National Rice Research Institute	春江157 Chunjiang157^、中嘉8号Zhongjia8hao^、春江糯8号Chunjiangnuo8、春江糯7号 Chunjiangnuo7、春江195 Chunjiang195、春江191 Chunjiang191、春江190 Chunjiang190、春江188 Chunjiang188、春江183 Chunjiang183、春江171 Chunjiang171
浙江省农业科学院 Zhejiang Academy of Agricultural Sciences	浙糯106 Zhenuo106^、浙粳96 Zhejing96^、ZH19-6、ZH19-12、ZH19-02、ZH18-47、ZH18-43、ZH18-117、ZH17-122、ZH16-98、R9079、R202、R201、R192、R1825、R182、R1751
宁波市农业科学研究院 Ningbo Academy of Agricultural Sciences	宁84 Ning84^,宁88 Ning88^、宁81 Ning81^、宁82 Ning82^、宁39 Ning39,甬粳19-46 Yongjing19-46、甬粳18-60 Yongjing18-60
嘉兴市农业科学研究院 Jiaxing Academy of Agricultural Sciences	秀水14 Xiushui14^、秀水134 Xiushui134^、秀水121 Xiushui121^、嘉67 Jia67^、天微香粳 Tianweijing、嘉禾628 Jiahe218、嘉60 Jia60、嘉16-48 Jia16-48、嘉124 Jia124、丙19-339 Bing19-339、丙19-26 Bing19-26、丙18-979 Bing18-979、丙18-927 Bing18-927、丙17-204 Bing17-204、丙17-02 Bing17-02、丙16-135 Bing16-135、丙16-111 Bing16-111、丙10-544 Bing10-544
湖州市农业科学研究院 Huzhou Academy of Agricultural Sciences	HZ16-78、HZ19-1137B、HZ19-1137C
绍兴市农业科学研究院 Shaoxing Academy of Agricultural Sciences	绍糯9714 Shaonuo9714^、绍粳18 Shaojing18^、绍粳19-6 Shaojing19-6 、绍粳19-34 Shaojing19-34、绍粳19-19 Shaojing19-19
台州市农业科学研究院 Taizhou Academy of AgriculturalSciences	台19-84 Tai19-84

目的基因 Target genes	引物名称 Primer name	序列(5'-3') Sequence (5'-3')	片段大小 Fragment size/bp
Pita^[16]	YL155/YL87-F	AGCAGGTTATAAGCTAGGCC	1042(R)
	YL155/YL87-R	CTACCAACAAGTTCATCAAA
Pib^[17]	PiBdom-F	GAACAATGCCCAAACTTGAGA	365(R)
	PiBdom-R	GGGTCCACATGTCAGTGAGC
Pigm	M143104-F	CCTTGTTCCTCCTGCTATC	215(R)/288(S)
	M143104-R	ATCTCGCTGTTCAGTCTTG
Pikh/Pi54^[18]	Pi54-F	CAATCTCCAAAGTTTTCAGG	216(R)/359(S)
	Pi54-R	GCTTCAATCACTGCTAGACC
Pi9^[19]	Pi9-F	GCTGTGCTCCAAATGAGGAT	291 (R)/397(S)
	Pi9-R	GCGATCTCACATCCTTTGCT
Pi1^[20]	M-Pi1-F	GTGCTGCTGTGGCTAGTTTG	460(R)
	M-Pi1-R	AGTCCCCGCTCAATTTTTCT
Pi2^[21]	Pi2CM1-F	CACCGTCGAACCACACCGTCAC	301(R)/352/363(S)
	Pi2CM1-R	ATCTGCACGACAGTGGCTGGTC
Pi5^[19]	Pi5-F	ATAGATCATGCGCCCTCTTG	206 (R)/307 (S)
	Pi5-R	TCATACCCCATTCGGTCATT
Pikm^[22]	Pik1-F	TCGAGTTGCTGGAACAAGGG	575(R)
	Pik1-R	ACTGCGTTCCCAATGCATGA
Pit^[23]	tRn1-F	GTTGGAGCTACGGTTGTTCAG	733(R)
	tRn1-R	ATGATAACCTCATCCTCAATAAGT
Pb1^[24]	PB1-F	ATCAACGCTACCTTCCC	160(R)
	PB1-R	GTGCCATCACAATTTCTTC
Bph14^[25]	B14InD-F	CTACTGATTGCAGATTGACG	324(R)/440(S)
	B14InD-R	CACTTGGTGAACTTATTCCCTT
Bph15^[26]	B15InD1-F	TGCACCTGGATCAAATGTTTCT	460(R)/360(S)
	B15InD1-R	GCCCTTCCCTCTCCATATCC
Xa21^[27]	U1-F	CGATCGGTATAACAGCAAAA	574(R)/447(S)
	I2-R	TCTGATCATGCATGTTCTGTG
Xa23^[28]	Xa23FunF	AAAGTCCCTTCCGAAACATC	105(R)
	Xa23FunR	ATGAGGAAGTGCTGCCAGA

浙江近年育成粳稻新品种(系)部分抗病虫基因的分子检测与育种应用

Molecular detection and breeding application of some disease and insect resistance genes of japonica rice varieties/lines recently developed in Zhejiang Province

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品种(系) Varieties/lines	抗性基因Resistance genes
品种(系) Varieties/lines	Pi2	Pigm	Pi9	Pita	Pib	Pi54/Pish	Pit	Pb1	Pi5	Pikm	Bph14	Bph15
甬粳19-46 Yongjing19-46	+	-	-	-	-	-	-	-	-	+	-	-
甬粳18-60 Yongjing18-60	+	-	-	-	+	+	+	-	-	-	-	-
中嘉8号 Zhongjia8	-	-	+	+	+	-	-	-	-	-	-	-
天微香粳 Tianweijing	+	-	-	-	+	-	-	-	+	+	-	-
台19-84 Tai19-84	+	-	-	-	+	-	-	-	-	-	-	-
绍糯9714 Shaonuo9714	-	-	+	+	+	+	-	-	+	-	-	-
品种(系) Varieties/lines	抗性基因Resistance genes
品种(系) Varieties/lines	Pi2	Pigm	Pi9	Pita	Pib	Pi54/Pish	Pit	Pb1	Pi5	Pikm	Bph14	Bph15
绍粳19-6 Shaojing19-6	+	-	-	-	+	+	-	-	+	-	-	-
绍粳19-34 Shaojing19-34	-	-	+	-	+	-	-	-	-	-	-	-
绍粳19-19 Shaojing19-19	+	-	-	+	+	-	-	-	+	-	-	-
绍粳18 Shaojing18	-	-	+	-	+	+	-	-	-	+	-	-
宁88 Ning88	+	-	-	+	+	+	-	-	-	-	-	-
宁84 Ning84	+	-	-	-	+	+	-	-	-	-	-	-
宁82 Ning82	+	-	-	+	+	-	-	-	-	-	-	-
宁81 Ning81	-	-	+	-	+	+	-	-	-	+	-	-
宁39 Ning39	+	-	-	+	+	+	-	-	-	+	-	-
嘉禾628 Jiahe628	-	-	+	-	+	-	-	-	+	+	-	-
嘉67 Jia67	+	-	-	-	+	-	+	-	-	-	-	-
嘉60 Jia60	-	-	-	-	+	-	-	-	+	-	-	-
嘉16-48 Jia16-48	-	-	-	-	+	-	-	-	+	+	-	-
嘉124 Jia124	-	-	+	-	+	-	-	-	+	+	-	-
春江糯8号 Chunjiangnuo8	-	-	+	+	+	+	-	-	+	+	-	-
春江糯7号Chunjiangnuo7	+	-	-	+	+	-	-	-	+	+	-	-
春江195 Chunjiang195	+	-	-	+	+	-	-	-	+	+	-	-
春江191 Chunjiang191	+	-	-	-	+	-	-	-	+	+	-	-
春江190 Chunjiang190	+	-	-	-	+	-	-	-	-	+	-	-
春江188 Chunjiang188	+	-	-	-	+	-	-	+	+	+	-	+
春江183 Chunjiang183	+	-	-	-	+	+	-	+	+	+	-	-
春江171 Chunjiang171	+	-	-	+	+	-	-	-	-	+	-	-
春江157 Chunjiang157	+	-	-	+	+	-	+	-	-	+	-	-
秀水14 Xiushui14	-	-	+	+	+	-	+	-	-	+	-	-
秀水134 Xiushui134	+	-	-	+	+	-	-	-	-	+	-	-
秀水121 Xiushui121	+	-	-	+	+	-	+	-	-	+	-	-
丙19-339 Bing19-339	-	-	-	+	-	-	-	-	-	+	-	+
丙19-26 Bing19-26	+	-	-	+	-	-	+	-	-	+	+	+
丙18-979 Bing18-979	+	-	-	+	+	-	+	-	-	-	-	-
丙18-927 Bing18-927	+	-	-	+	+	-	-	-	-	-	-	-
丙17-204 Bing17-204	+	-	-	+	+	+	+	-	-	+	-	+
丙17-02 Bing17-02	+	-	-	+	+	-	+	-	+	+	+	+
丙16-135 Bing16-135	+	-	-	+	+	-	+	-	-	+	+	+
丙16-111 Bing16-111	+	-	-	+	+	-	+	+	-	+	+	+
丙10-544 Bing10-544	+	-	-	-	+	-	+	+	-	-	-	-
浙粳96 Zhejing96	+	-	-	+	+	-	+	-	-	+	-	-
ZH19-6	+	-	-	+	+	-	-	-	-	+	-	-
ZH19-12	+	-	-	+	-	-	+	-	-	-	-	-
ZH19-02	-	-	+	+	+	-	+	-	-	+	-	-
ZH18-47	+	-	-	+	+	-	-	-	-	+	-	-
ZH18-43	+	-	-	+	+	+	-	-	-	+	-	-
ZH18-117	+	-	-	+	+	+	+	-	-	+	-	-
ZH17-122	+	-	-	+	+	-	+	-	-	+	-	-
ZH16-98	+	-	-	+	+	±	-	-	-	+	-	-
浙糯106 Zhenuo106	+	-	-	-	+	-	-	-	+	+	-	-
R9079	+	-	-	+	+	-	-	-	-	+	-	-
R202	-	-	+	+	+	-	-	-	-	+	-	-
R201	+	-	-	+	+	-	-	-	-	+	-	-
R192	-	-	+	+	+	-	+	-	+	+	-	-
R1825	-	+	-	+	+	+	-	-	-	+	-	-
R182	-	-	+	+	+	-	+	-	+	+	-	-
R1751	-	-	+	-	+	-	+	-	-	-	-	-
HZ19-1137C	-	-	+	-	+	-	-	-	+	-	-	-
HZ19-1137B	-	-	+	+	+	-	-	-	+	-	-	-
HZ16-78	+	-	-	+	-	-	+	-	-	+	-	-
检出率Detection rate/%	67.21	1.64	26.23	63.93	91.80	24.59	34.43	6.56	32.79	68.85	6.56	11.48

品名(系) Varieties/lines	来源 Origins	Bph14	Bph15	抗性均值 Mean resistance	抗性级别 Resistance grade	抗性水平 Resistance level
B5	—	+	+	1.86	1	HR
TN1	—	-	-	8.16	9	HS
宁84 Ning84	晚粳品种Late japonica rice	-	-	6.73	7	S
宁88 Ning88	晚粳品种Late japonica rice	-	-	7.03	7	S
H1299	F₅(宁84×丙16-111)(Ning 84×Bing 16-111)	+	+	3.96	3	R
H241	F₅(丙16-111×宁84)(Bing 16-111×Ning 84)	+	+	3.55	3	R
H234	F₅(宁84×丙16-111)(Ning 84×Bing 16-111)	-	+	4.43	5	MR
H6050	F₅(宁88×丙16-111)(Ning 84×Bing 16-111)	-	+	5.02	5	MR
H6034	F₅(宁88×丙16-111)(Ning 84×Bing 16-111)	+	+	3.23	3	R
H6086	F₅(宁88×丙16-111)(Ning 84×Bing 16-111)	+	+	3.35	3	R

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