用SRAP标记研究根际土壤微生物的遗传多样性

doi:10.3724/SP.J.1003.2011.09232

摘要/Abstract

摘要：

将近年来新建立的分子标记技术——相关序列扩增多态性(sequence-related amplified polymorphism, SRAP)应用于土壤微生物遗传多样性研究。采用22对引物组合对20种植物根际土壤微生物进行了分析, 共获得237个扩增位点, 其中多态性位点221个, 占93.2%。平均每对引物组合的多态位点比例(PPL)、多态信息含量(PIC)、等位基因单体型(Ah)和遗传杂合度(He)分别为93.78%、0.94、18.05和0.92, 说明SRAP对根际土壤微生物具有较强的鉴别能力, 也反映了本研究中20种不同植物的根际土壤微生物具有丰富的遗传多样性。水稻的2个不同种植地和4个不同发育时期间的根际土壤微生物遗传距离差异极显著, 但2个不同水稻品种间的差异不显著。Shannon多样性指数揭示, 水稻根际土壤微生物的遗传多样性最低, 莴苣的最高。按照非加权类平均法(UPGMA)聚类, 在遗传距离0.454的水平上, 可将20种植物根际土壤微生物分为三类: 第一类是水稻根际土壤微生物, 第二类是种植于大棚温室的芹菜根际土壤微生物, 第三类为其余18种旱作植物根际土壤微生物。本研究结果证明SRAP是分析土壤微生物遗传多样性的有效手段。

关键词: 根际土壤微生物, SRAP, 遗传多样性, 遗传距离, 聚类分析

Abstract

We have attempted to use the SRAP (sequence-related amplified polymorphism) markers, a new molecular technology, to study genetic diversity in soil microbes. We sampled rhizosphere soil microbes from 20 plant species and employed 22 SRAP primer combinations. A total of 237 scorable fragments were identified, of which 221 (93.2%) were polymorphic loci. The average percentage of polymorphic loci (PPL), polymorphism information content (PIC), allele haplotype (Ah), and expected heterozygosity (He) for each primer combination were 93.78%, 0.94, 18.05 and 0.92, respectively. Our results revealed rich genetic diversity in rhizosphere soil microbes and the high ability of SRAP to resolve samples based on their genetic basis. Differences in genetic distance for rice rhizosphere microbes between two locations, and that among four different developmental stages were both significant at 0.01 level, but the difference was not significant between two varieties. Shannon diversity indices indicated that the genetic diversity of rhizosphere soil microbes was lowest in rice and highest in lettuce. The rhizosphere soil microbes from 20 plant species could be clustered into three groups at the 0.454 (GD) level based on UPGMA, in which the first group was from rice, while the second group was from celery planted in plastic green house, and the third group was from 18 other plant species cultivated in dry lands. Our results suggest that SRAP is an efficient method for analyzing the genetic diversity in rhizosphere soil microbes.

Key words: rhizosphere soil microbe, SRAP, genetic diversity, genetic distance, cluster analysis

李春楠, 崔海瑞, 王伟博 (2011) 用SRAP标记研究根际土壤微生物的遗传多样性. 生物多样性, 19, 485-493. DOI: 10.3724/SP.J.1003.2011.09232.

Chunnan Li, Hairui Cui, Weibo Wang (2011) Genetic diversity in rhizosphere soil microbes detected with SRAP markers. Biodiversity Science, 19, 485-493. DOI: 10.3724/SP.J.1003.2011.09232.

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链接本文: https://www.biodiversity-science.net/CN/10.3724/SP.J.1003.2011.09232

https://www.biodiversity-science.net/CN/Y2011/V19/I4/485

图/表 11

参考文献 32

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编号 Code	植物 Plant species
A 区 Zone A
OS	水稻 Rice (Oryza sativa)
B 区 Zone B
AG	芹菜 Celery (Apium graveolens)
AS	大蒜 Garlic (Allium sativum)
AT	韭菜 Chinese chives (Allium tuberosum)
BC	白菜 Chinese cabbage (Brassica campestris ssp. pekinensis)
BN	油菜 Rapeseed (Brassica napus)
CS	茶树 Tea plant (Camellia sinensis)
GM	大豆 Soybean (Glycine max)
LE	番茄 Tomato (Lycopersicon esculentum)
LS	莴苣 Lettuce (Lactuca sativa)
PS	豌豆 Pea (Pisum sativum)
RS	萝卜 Radish (Raphanus sativus)
SM	茄子 Eggplant (Solanum melongena)
ST	马铃薯 Potato (Solanum tuberosum)
TA	普通小麦 Wheat (Triticum aestivum)
ZM	玉米 Maize (Zea mays)
C 区 Zone C
BO	甘蓝 Cabbage (Brassica oleracea)
FA	高羊茅 Tall fescue (Festuca arundinacea)
HV	大麦 Barley (Hordeum vulgare)
VF	蚕豆 Broad bean (Vicia faba)

编号 Code	植物 Plant species
A 区 Zone A
OS	水稻 Rice (Oryza sativa)
B 区 Zone B
AG	芹菜 Celery (Apium graveolens)
AS	大蒜 Garlic (Allium sativum)
AT	韭菜 Chinese chives (Allium tuberosum)
BC	白菜 Chinese cabbage (Brassica campestris ssp. pekinensis)
BN	油菜 Rapeseed (Brassica napus)
CS	茶树 Tea plant (Camellia sinensis)
GM	大豆 Soybean (Glycine max)
LE	番茄 Tomato (Lycopersicon esculentum)
LS	莴苣 Lettuce (Lactuca sativa)
PS	豌豆 Pea (Pisum sativum)
RS	萝卜 Radish (Raphanus sativus)
SM	茄子 Eggplant (Solanum melongena)
ST	马铃薯 Potato (Solanum tuberosum)
TA	普通小麦 Wheat (Triticum aestivum)
ZM	玉米 Maize (Zea mays)
C 区 Zone C
BO	甘蓝 Cabbage (Brassica oleracea)
FA	高羊茅 Tall fescue (Festuca arundinacea)
HV	大麦 Barley (Hordeum vulgare)
VF	蚕豆 Broad bean (Vicia faba)

取样地点 Sites	pH	有机质 Organic matter (g/kg)	全氮 Total N (g/kg)	速效P Available phosphorus (mg/kg)	速效K Available potassium (mg/kg)
A区	6.47	18.59	1.13	18.40	43.80
B区	7.20	16.70	1.44	52.54	61.76
C区	7.15	15.10	1.80	48.52	58.10

取样地点 Sites	pH	有机质 Organic matter (g/kg)	全氮 Total N (g/kg)	速效P Available phosphorus (mg/kg)	速效K Available potassium (mg/kg)
A区	6.47	18.59	1.13	18.40	43.80
B区	7.20	16.70	1.44	52.54	61.76
C区	7.15	15.10	1.80	48.52	58.10

编号 Code	正向引物序列 Forward primer (5'-3')	编号 Code	反向引物序列 Reverse primer (5′-3′)
F1	TGAGTCCAAACCGGATA	R1	GACTGCGTACGAATTAAT
F2	TGAGTCCAAACCGGAGC	R2	GACTGCGTACGAATTTGC
F3	TGAGTCCAAACCGGAAT	R3	GACTGCGTACGAATTGAC
F4	TGAGTCCAAACCGGACC	R4	GACTGCGTACGAATTTGA
F5	TGAGTCCAAACCGGAAG	R5	GACTGCGTACGAATTAAC
F6	TGAGTCCAAACCGGTAA	R6	GACTGCGTACGAATTGCA
F7	TGAGTCCAAACCGGTCC	R7	GACTGCGTACGAATTCAA
F8	TGAGTCCAAACCGGTGC	R8	GACTGCGTACGAATTAGC