金佛山方竹林AM真菌调查及接种对其苗期生长的影响

接种AM真菌对金佛山方竹生长的影响

摘要

AM真菌是一种专性活体营养型真菌，通过与植物建立共生体，促进生长发育、提高产量、改善品质，被作为新型生物肥料广泛应用于农林及园艺等领域。论文采集金佛山方竹天然林根际土壤，测定根系侵染状况，应用形态学和分子生物学方法鉴定AM真菌主要种类和优势种属。以三叶草为宿主植物，扩繁常见AM真菌（根内根生囊霉Rhizophagus intraradices和摩西管孢囊霉Funneliformis mosseae）和金佛山方竹根际土壤优势AM真菌（蜜色无梗囊霉Acaulospora mellea 和美丽盾巨孢囊霉Scutellospora calospora），以扩繁菌剂接种金佛山方竹实生苗，测定金佛山方竹不同时期生物量、侵染率、光合作用、酶活性等指标，研究接种AM真菌对金佛山方竹生长发育的影响，试验结果如下：

1、在金佛山方竹天然林分布区域的贵州省正安县新州镇、桐梓县狮溪镇采集12份根系和根际土壤，碱解离-酸性品红染色法测定了AM真菌根系侵染率，根系侵染率均超过30%，镜检观察到大量AM真菌菌丝、泡囊、丛枝等结构，说明金佛山方竹能与AM真菌建立良好的共生关系。

2、采用湿筛倾析-蔗糖离心法对金佛山方竹根际土壤AM真菌孢子进行分离，通过形态鉴定，鉴定出7属24种，新增6属15种。其中，球囊霉属Glomus 7种，无梗囊霉属Acaulospora8种，硬内囊霉属Sclerocystis1种，盾巨孢囊霉Scutellospora3种，巨孢囊霉属Gigaspora3种，盾孢囊霉属Racocetra1种，伞房球囊霉Corymbiglomus1种。采用nested-PCR技术对AM真菌18S rDNA AML1-AML2区序列进行系统发育分析，以6属15种AM真菌为原材料进行分子鉴定，鉴定出2种AM真菌为球囊霉科Glomeraceae。

3、接种4种AM真菌均能促进金佛山方竹实生苗的生长，促生效果因AM真菌种类而异。试验结果显示：在幼苗长出第三片真叶时接种美丽盾巨孢囊霉Scutellospora calospora侵染效果最好，侵染率达到60.49%；种子发芽期接种摩西管孢囊霉Funneliformis mosseae对金佛山方竹生物量特别是鲜重的影响最显著，比不接种处理提高了145.71%；在种子萌发期接种摩西管饱囊霉Funneliformis mosseae对金佛山方竹净光合速率和蒸腾速率的影响存在显著性差异，比不接种分别提高了276.89%和59.46%，而其叶绿素含量比不接种处理提高53.36%；在

幼苗第三真叶期接种美丽盾巨孢囊霉Scutellospora calospora对金佛山方竹POD活性的影响差异显著，比不接种处理显著提高了77.91%；在幼苗第一真叶期接种根内根生囊霉Rhizophagus intraradices对金佛山方竹SOD活性的影响存在显著性差异，比不接种处理显著提高了98.21%；在种子萌发期接种摩西管饱囊霉Funneliformis mosseae对硝酸还原酶活性差异最显著，比不接种处理显著提高了145.00%；在幼苗第一真叶期接种美丽盾巨孢囊霉Scutellospora calospora 的可溶性糖含量比不接种处理显著提高了58.50%。就接种不同AM真菌种类而言，摩西管孢囊霉Funneliformis mosseae和美丽盾巨孢囊霉Scutellospora calospora的促生效应最显著，在金佛山方竹人工林生产中，可作为适宜接种的AM真菌菌株。

试验结果明确了金佛山方竹天然林根际土壤中AM真菌主要种类和优势种，筛选了适宜接种的AM真菌菌株，这将为AM真菌在金佛山方竹人工林生产中的应用提供种质资源、技术支持和理论依据。

关键词：金佛山方竹；AM真菌；菌根效应；nested-PCR技术；酶活性；生物量

Effects of AM fungi on the growth of

Chimonobambusa utilis

Abstract

Arbuscular mycorrhizal(AM)fungi was a kind of obligate viviparous fungi and can form a symbiotic association with host plant.This kind of symbiosis can promote plant growth,improve crop yields and qualities,and improved soil structure,so the m ycorrhizal bio-fertilizers has gradually gotten popularized in the domain of agriculture and forestry.In this paper,the soil that the natural forest rhizosphere of Chimonoba mbusa utilis was collected,and the infection status of root system was determined.Th e main species and dominant species of AM fungi were identified by morphological a nd molecular biological methods.The effect that AM fungi on the growth of different growth period by inoculated with AM fungi of autochthonic(Rhizophagus intraradice s and Funneliformis mosseae)and dominant of roots(Acaulospora mellea and Scutell ospora calospora),to which gain the fine mycorrhizal.The mian results are as follows:

1.Different mycorrhizal structure in all root sample was observed under microsc ope and the infection rates of Zhenan county and Tongzi county of Guizhou were hig her than30%with used alkaline lysis and acid fuchsin staining method.The results sh owed Chimonobambusa utilis can establish a good symbiosis relation with AM fungi.

2.There are15new species of six genera that twenty-four species of AM fungi b elonged to seven genera with based on the morphological characteristics of the isolate d spores from rhizoshere soil of Chimonobambusa utilis,which including seven speci es of Glomus,eight species of Acaulospor a,one species of Sclerocystis,three species of Scutellospora,three species of Gigaspora,one species of Corymbiglomus.Phyloge netic analysis of the18S rDNA AML1-AML2sequence was used to identify the AM fungi of15new species by nested-PCR,only two of which were consistent with morp hological identification as Glomeraceae.

3.Inoculation of4kinds of AM fungi can promote the growth of Chimonobambu sa utilis,but the effect was different because of the type of AM fungi.The result sho ws the best infection rate was Scutellospora calospora inoculated with the third leaf