不同基因型腐烂茎线虫群体杂交后代分子特征比较

doi:10.13560/j.cnki.biotech.bull.1985.2021-0576

摘要/Abstract

摘要：

为了明确不同基因型腐烂茎线虫（Ditylenchus destructor）是否具有生殖隔离及杂交后代的变异程度,选用1条3-4龄幼虫与5条雄虫,将不同基因型群体进行生物学杂交,对F1代数量、ITS-rDNA序列及其二级结构比对分析。结果表明,杂交产生F1代数量普遍较少,不同基因型杂交组合F1代线虫数量≤6条/处理,部分组合的后代仅为1条。因F1代数量较少,11个杂交组合中获得5个F1代群体的ITS-rDNA序列,不同基因型杂交F1代与其亲本ITS-rDNA序列均存在0-86 bp的碱基差异,且其二级结构与亲本也有差异,核糖体ITS-RFLP结果呈现丰富的多态性,系统发育分析发现部分F1代群体与父本亲缘关系较近,部分与母本较近。不同基因型群体未发现生殖隔离,杂交F1代的变异主要出现在ITS1区H9螺旋区域,且变异程度与亲本间的差异呈正比。本研究通过模拟试验分析了腐烂茎线虫不同群体扩散后可能产生的杂交代基因型分化,研究结果对于腐烂茎线虫遗传多样性研究和防治实践均具有参考价值。

关键词: 腐烂茎线虫, 基因型, 杂交, RNA二级结构

Abstract:

In order to clarify whether there are reproductive isolation and the variation in the hybrid offspring between different haplotypes,1 of 3^rd to 4^th stage juveniles and 5 males from different haplotypes were selected to hybridize,and the nematode amounts,ITS-rDNA sequences and secondary structures of F1 generations were analyzed. The results showed that the nematode amount of F1 generations reproduced after hybridized were generally low,and F1 amount of the different cross groups were ≤ 6 per treatment. In some groups,only one nematode was found in one treatment. Due to the low amount of offspring,the ITS-rDNA sequences of 5 F1 generations among 11 groups were successfully sequenced. The ITS-rDNA sequence of F1 generations were different with 0-86 bp from their parents,and all of the secondary structures of ITS were different from those of the parents. The results of ribosomal ITS-RFLP showed diversiform. Phylogenetic analysis showed that some of the F1 generation groups were closer to the male parents,and some were closer to the female parents. No reproductive isolation was found in different haplotypes,but variation appeared mainly in the H9 helix region of ITS1 region in the F1 generation of hybrids,and the degree of variation was directly proportional to the differences between parents. The analysis of differentiation on hybrid generations from different haplotypes of D. destructor were conducted through simulation tests after population diffusions. The results may provide references both to further researches on genetic diversity and control practice of D. destructor.

Key words: Ditylenchus destructor, haplotype, hybridization, RNA secondary structure

倪春辉, 李惠霞, 李文豪, 刘永刚, 徐雪芬, 韩变. 不同基因型腐烂茎线虫群体杂交后代分子特征比较[J]. 生物技术通报, 2021, 37(7): 118-126.

NI Chun-hui, LI Hui-xia, LI Wen-hao, LIU Yong-gang, XU Xue-fen, HAN Bian. Comparison of Molecular Characteristics of the Hybrid Progenies from Different Haplotypes of Ditylenchus destructor[J]. Biotechnology Bulletin, 2021, 37(7): 118-126.

图/表 8

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序号 Serial number	线虫编号 Sample No.	寄主 Host	采集地 Collecting location	基因型 Haplotype
1	DXP1	马铃薯	甘肃省定西市安定区	B
2	DXP2	马铃薯	甘肃省定西市安定区	C
3	HLJP1	马铃薯	黑龙江绥化市	C
4	MXA2	当归	甘肃省定西市岷县	N
5	TCC1	党参	甘肃省定西市宕昌县	B
6	ZXA1	当归	甘肃省定西市漳县	H
7	WYA4	党参	甘肃省定西市渭源县	K

序号 Serial number	线虫编号 Sample No.	寄主 Host	采集地 Collecting location	基因型 Haplotype
1	DXP1	马铃薯	甘肃省定西市安定区	B
2	DXP2	马铃薯	甘肃省定西市安定区	C
3	HLJP1	马铃薯	黑龙江绥化市	C
4	MXA2	当归	甘肃省定西市岷县	N
5	TCC1	党参	甘肃省定西市宕昌县	B
6	ZXA1	当归	甘肃省定西市漳县	H
7	WYA4	党参	甘肃省定西市渭源县	K

亲本（Parent）♀	亲本（Parent）♂
亲本（Parent）♀	DXP1	DXP2	MXA2	TCC1	ZXA1	WYA4
DXP1	⊕	×	Φ	×	Φ	×
DXP2	×	Φ	×	Φ	Φ	Φ
MXA2	×	×	Φ	Φ	×	Φ
TCC1	Φ	Φ	Φ	Φ	Φ	Φ
ZXA1	Φ	Φ	×	Φ	Φ	Φ
HLJP1	×	Φ	Φ	Φ	Φ	Φ

亲本（Parent）♀	亲本（Parent）♂
亲本（Parent）♀	DXP1	DXP2	MXA2	TCC1	ZXA1	WYA4
DXP1	⊕	×	Φ	×	Φ	×
DXP2	×	Φ	×	Φ	Φ	Φ
MXA2	×	×	Φ	Φ	×	Φ
TCC1	Φ	Φ	Φ	Φ	Φ	Φ
ZXA1	Φ	Φ	×	Φ	Φ	Φ
HLJP1	×	Φ	Φ	Φ	Φ	Φ

杂交组合（父本×母本） Hybridized combination （Male parent× Female parent）	重复管数 Number of repetitions	产生后代杂交管数 Number of progeny-produced tubes	平均后代数（条/管） Average number of offspring （lines per tube）
DXP1×DXP1	20	5	18±2.7b
DXP2×DXP1	20	5	6±1.7b
DXP1×DXP2	20	3	2±0.6b
WYA4×DXP1	20	2	1±0b
DXP1× HLJP1	20	1	1±0b
MXA2×ZXA1	20	5	1±0b
ZXA1×MXA2	20	4	4±1.0b
MXA2×DXP2	20	2	1±0b
DXP2×MXA2	20	3	1±0b
TCC1×DXP1	20	6	103±79.5a
TCC1×HLJP1	20	2	3±1.0b