转基因玉米连续种植对土壤丛枝菌根真菌群落的影响

doi:10.11924/j.issn.1000-6850.casb2024-0062

中国农学通报 ›› 2024, Vol. 40 ›› Issue (29): 65-74.doi: 10.11924/j.issn.1000-6850.casb2024-0062

• 资源·环境·生态·土壤·气象 • 上一篇下一篇

转基因玉米连续种植对土壤丛枝菌根真菌群落的影响

刘仟龙¹(), 刘瑞华¹^,², 李刚¹, 修伟明¹, 杨殿林¹, 刘红梅¹(), 赵建宁¹()

¹ 农业农村部环境保护科研监测所，天津 300191
² 中国农业科学院棉花研究所，河南安阳 455000

收稿日期:2024-01-12 修回日期:2024-07-21 出版日期:2024-10-15 发布日期:2024-10-14
通讯作者:
刘红梅，女，1976年出生，河北沧州人，高级工程师，博士，主要从事转基因作物土壤环境安全评价方面的研究。通信地址：300191 天津市南开区复康路31号农业农村部环境保护科研监测所，Tel：022-23611820，E-mail：liuhongmei@caas.cn；
赵建宁，男，1981年出生，陕西西安人，研究员，博士，主要从事生物多样性与生态农业研究。通信地址：300191 天津市南开区复康路31号农业农村部环境保护科研监测所，Tel：022-23611820， E-mail：zhaojianning@caas.cn。
作者简介:
刘仟龙，男，2000年出生，湖南邵阳人，硕士研究生，研究方向：农业面源污染与生态治理。通信地址：300191 天津市南开区复康路31号农业农村部环境保护科研监测所，Tel：022-23611820，E-mail：806621205@qq.com。
基金资助:
“国家转基因重大专项”(2016ZX08012005-005)

Effects of Continuous Planting of Transgenic Maize on Community of Arbuscular Mycorrhizal Fungi in Soil

LIU Qianlong¹(), LIU Ruihua¹^,², LI Gang¹, XIU Weiming¹, YANG Dianlin¹, LIU Hongmei¹(), ZHAO Jianning¹()

¹ Agro-environmental Protection Institute, Ministry of Agriculture and Rural Affairs, Tianjin 300191
² Institute of Cotton Research of Chinese Academy of Agricultural Sciences, Anyang, Henan 455000

Received:2024-01-12 Revised:2024-07-21 Published:2024-10-15 Online:2024-10-14

摘要/Abstract

摘要：

为评估转基因玉米连续种植对土壤丛枝菌根真菌群落结构的影响，以转基因玉米DBN9936和其受体玉米DBN318为材料，采用PCR-DGGE技术和PLFA技术分析土壤AM真菌群落结构特征。结果表明：在相同的种植年份和地点下，转基因和非转基因玉米的土壤AM真菌群落结构表现出较高的相似性，其香农-威纳指数、Pielou均匀度指数和丰富度均未出现显著性差异，且优势属均为Clomus（球囊霉属）。聚类分析结果显示，同一种植地点下的2种玉米土壤中AM真菌群落的相似度大多在0.60以上，而不同地点的群落相似度均小于0.60；同时，不同种植地点的两种玉米非常清晰的分布在聚类图的上下两侧，说明土壤AM真菌群落结构受种植地点影响较大，而转基因本身对其影响较为微弱。系统发育分析结果表明，不同地点下种植的两种玉米的特有条带在系统发育树上聚类在不同的类群。土壤微生物PLFA含量结果显示，转基因玉米DBN9936的种植对土壤微生物的总PLFA含量均未产生显著影响，各微生物类群的相对丰度差异也较小；而在不同种植地点下两种玉米土壤微生物的总PLFA含量均有显著性差异，且各微生物类群的相对丰度差异较大。综上，转基因玉米DBN9936较之对应的非转基因玉米对土壤AM真菌群落结构和PLFA含量无显著影响，但不同种植地点之间存在差异。

关键词: 转基因玉米, 丛枝菌根真菌, PCR-DGGE, PLFA, 群落结构

Abstract:

To evaluate the effect of continuous planting of transgenic maize on the community structure of soil arbuscular mycorrhizal fungi, transgenic maize DBN9936 and its non-transgenic counterpart were used as the experimental materials, the characteristics of soil arbuscular mycorrhizal (AM) fungal community structure were analyzed using PCR-DGGE and PLFA techniques. The results indicated that there was a high similarity in the soil AM fungal community structure between transgenic and non-transgenic maize in the same planting year and location. There were no significant differences in the Shannon-Wiener index, Pielou evenness index and richness. The dominant genera consistently identified were Glomus. Cluster analysis revealed that the similarity of AM fungal communities in soils from the two maize types at the same planting location was mostly above 0.60, while the similarity between communities from different locations was below 0.60. Additionally, the clustering pattern clearly separated the two maize types from different planting locations on the upper and lower sides of the cluster diagram, indicated that the community structure of soil AM fungi was greatly affected by planting sites, while the influence of the transgenic itself was weak. Phylogenetic analysis also indicated that the specific bands of the two maize species planted in different locations clustered in different groups on the phylogenetic trees. The results of soil microbial PLFA content showed that the planting of transgenic maize DBN9936 had no significant effect on the total soil microbial PLFA content, and the differences in the relative abundance of microbial groups were small. In contrast, significant differences were observed in the total PLFA content of soil microbes between the two maize types planted at different locations, with substantial variations in the relative abundance of microbial groups. In conclusion, compared to non-transgenic maize, transgenic maize DBN9936 had no significant impact on the structure of soil AM fungal communities and PLFA content. However, notable differences were observed among different planting locations.

Key words: transgenic maize, arbuscular mycorrhizal fungi, PCR-DGGE, PLFA, community structure

刘仟龙, 刘瑞华, 李刚, 修伟明, 杨殿林, 刘红梅, 赵建宁. 转基因玉米连续种植对土壤丛枝菌根真菌群落的影响[J]. 中国农学通报, 2024, 40(29): 65-74.

LIU Qianlong, LIU Ruihua, LI Gang, XIU Weiming, YANG Dianlin, LIU Hongmei, ZHAO Jianning. Effects of Continuous Planting of Transgenic Maize on Community of Arbuscular Mycorrhizal Fungi in Soil[J]. Chinese Agricultural Science Bulletin, 2024, 40(29): 65-74.

图/表 12

参考文献 37

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试验地	全磷/(g/kg)	全氮/(g/kg)	土壤有机质/(g/kg)	pH
吉林	0.52	1.02	9.35	5.16
河北	0.53	0.49	9.35	8.37

试验地	全磷/(g/kg)	全氮/(g/kg)	土壤有机质/(g/kg)	pH
吉林	0.52	1.02	9.35	5.16
河北	0.53	0.49	9.35	8.37

巢式PCR	引物	引物序列(5′-3′)	反应条件
第一步	AML1	ATCAACTTTCGATGGTAGGATAGA	94℃ 3 min；94℃ 30 s，50℃ 30 s， 72℃ 45 s，35个循环；72℃ 7 min
第一步	AML2	GAACCCAAACACTTTGGTTTCC	94℃ 3 min；94℃ 30 s，50℃ 30 s， 72℃ 45 s，35个循环；72℃ 7 min
第二步	NS31	GC-CGCCCGCCGCGCGCGGCGGGCGGGGCGGGGGCA CGGGGGGTTGGA	94℃ 3 min；94℃ 30 s，50℃ 30 s， 72℃ 45 s，35个循环；72℃ 7 min
第二步	G101	GGGCAA GTCTGGTGCC GCC TGC TTT AAA CAC TCT A	94℃ 3 min；94℃ 30 s，50℃ 30 s， 72℃ 45 s，35个循环；72℃ 7 min

巢式PCR	引物	引物序列(5′-3′)	反应条件
第一步	AML1	ATCAACTTTCGATGGTAGGATAGA	94℃ 3 min；94℃ 30 s，50℃ 30 s， 72℃ 45 s，35个循环；72℃ 7 min
第一步	AML2	GAACCCAAACACTTTGGTTTCC	94℃ 3 min；94℃ 30 s，50℃ 30 s， 72℃ 45 s，35个循环；72℃ 7 min
第二步	NS31	GC-CGCCCGCCGCGCGCGGCGGGCGGGGCGGGGGCA CGGGGGGTTGGA	94℃ 3 min；94℃ 30 s，50℃ 30 s， 72℃ 45 s，35个循环；72℃ 7 min
第二步	G101	GGGCAA GTCTGGTGCC GCC TGC TTT AAA CAC TCT A	94℃ 3 min；94℃ 30 s，50℃ 30 s， 72℃ 45 s，35个循环；72℃ 7 min

微生物类群	磷脂脂肪酸标记
细菌	i14:0，14:0，i15:0，a15:0，15:0，i16:0，16:0，16:1ω7c，i17:0，a17:0，17:0，cy17:0，18:1ω7，i19:0，a19:0，19:0，cy19:0
革兰氏阳性细菌	i14:0，i15:0，a15:0，i16:0，i17:0，a17:0
革兰氏阴性细菌	16:1ω7c，16:1ω9c，cyc17:0，17:1ω8c，18:1ω7c，cyc19:0
真菌	16:1ω5c，18:1ω9c，18:2ω6c，18:3ω6c
AM真菌	16:1ω5c，18:1ω7
厌氧菌	A17:0，i17:0

转基因玉米连续种植对土壤丛枝菌根真菌群落的影响

Effects of Continuous Planting of Transgenic Maize on Community of Arbuscular Mycorrhizal Fungi in Soil

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条带编号	GeneBank 登录号	同源性最高的菌株	相似度/%
1	JQ218148.1	Uncultured Glomus clone	100
2	KX809136.1	Uncultured Rhizophagus clone	99
3	KY979384.1	Uncultured Glomus clone	100
6	KM365412.1	Uncultured Glomeraceae clone	100
7	KC579419.1	Uncultured Glomus clone	99
8	KY232438.1	Uncultured Glomus clone	99
9	LN906586.1	Uncultured Archaeospora clone	99
10	KU168035.1	Uncultured Claroideoglomu	100
12	LT856617.1	Uncultured Rhizophagus clone	99
13	LT856616.1	Uncultured Rhizophagus clone	100
14	EU332725.1	Uncultured Paraglomus clone	98
15	GQ140597.1	Uncultured Glomus clone	100
16	KC588997.1	Uncultured Glomus clone	98
17	JN559802.1	Uncultured Glomus clone	98
18	MG835506.1	Uncultured Glomus clone	99
19	KY979290.1	Uncultured Glomus clone	100
20	KM085113.1	Uncultured Archaeospora clone	97
21	KY979289.1	Uncultured Glomus clone	99
22	KY232529.1	Uncultured Glomus clone	99
23	HE613489.1	Uncultured Paraglomus clone	99
24	KU668988.1	Uncultured Claroideoglomu	99

条带编号	GeneBank 登录号	同源性最高的菌株	相似度/%
1	KY979378.1	Uncultured Glomus clone	100
2	KY232438.1	Uncultured Glomus clone	99
3	KY979384.1	Uncultured Glomus clone	100
4	KX154256.1	Uncultured Rhizophagus clone	100
5	KY173792.1	Uncultured Glomus clone	97
6	KY232420.1	Uncultured Glomus clone	98
7	LN621194.1	Uncultured Claroideoglomus	100
8	MG835539.1	Uncultured Rhizophagus clone	100
9	KU361755.1	Uncultured Glomus clone	99
10	KU668988.1	Uncultured Claroideoglomus	100
11	KY232471.1	Uncultured Glomus clone	100
12	MF567532.1	Uncultured Glomeromycotina clone	99
13	HG425740.1	Uncultured Glomus clone	100
14	KX462871.1	Uncultured Rhizophagus clone	99
15	KF601851.1	Uncultured Glomus clone	100
16	KT238942.1	Uncultured Glomeromycotina clone	100
17	KY979306.1	Uncultured Glomus clone	100
18	KY232617.1	Uncultured Glomus clone	99
19	MF567532.1	Uncultured Glomeromycotina clone	100
20	LT672514.1	Uncultured Claroideoglomus	99
21	KY232529.1	Uncultured Glomus clone	100

种植年份	种植地点	品种	香农-威纳指数(H')	丰富度(S)	均匀度指数(J)
2016年	河北省	DBN9936	2.94±0.12Aa	21.67±1.53Aa	0.96±0.02Aa
	河北省	DBN318	2.94±0.12Aa	21.67±1.53Aa	0.96±0.02Aa
	吉林省	DBN9936	2.83±0.14Aa	20.67±2.08Aa	0.94±0.02Aa
	吉林省	DBN318	2.83±0.14Aa	20.67±2.08Aa	0.94±0.02Aa
2017年	河北省	DBN9936	3.23±0.02Ab	29.33±0.58Ab	0.95±0.01Aa
	河北省	DBN318	3.10±0.11Ab	26.33±2.31Ab	0.95±0.01Aa
	吉林省	DBN9936	3.33±0.10Aa	31.67±2.08Aa	0.96±0.10Aa
	吉林省	DBN318	3.28±0.11Aa	30.33±2.31Aa	0.96±0.01Aa

种植地点	品种	2016年总PLFA含量/(nmol/g)	2017年总PLFA含量/(nmol/g)
河北省	DBN9936	45.19±2.93Ab	45.27±3.02Ab
河北省	DBN318	48.13±3.46Ab	44.13±4.01Ab
吉林省	DBN9936	55.59±3.44Aa	63.16±6.42Aa
吉林省	DBN318	59.42±3.65Aa	55.14±3.75Aa

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