内蒙古地区四种树木外生菌根形态多样性及分子鉴定
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摘要
本研究包括与不同树种共生的外生菌根类型的形态学、组织学特征分析及其分子鉴定研究两部分内容,现分述摘要如下:
(一)外生菌根形态学、解剖学特征分析
本研究从某个树种的角度出发,以菌根共生体为研究对象,系统地从形态学和解剖学的角度研究了与之共生的外生菌根类型,并以图谱及文字说明的形式对本研究实验中所采集到的菌根标本中的48种类型进行了详细描述;其中与青海云杉共生的外生菌根分出11种类型,与白桦共生的外生菌根分出13种类型,与山杨共生的外生菌根分出10种类型,与兴安落叶松共生的外生菌根14种类型。以宏观的形态、颜色和显微结构中菌套菌丝排列图式、菌套表面结构及菌丝和菌索内部结构等特征为主要依据进行外生菌根初步分类鉴定,并编制了检索表。
(二)外生菌根根尖分子鉴定研究
在依据形态学、解剖学特征分出基本类型的基础上,以各个树种的菌根共生体的根尖为材料,提取DNA,利用真菌特有引物ITS1F和ITS4,并在PCR基础上对外生菌根真菌rDNA的ITS区段进行碱基序列测定,然后与互联网上DNA序列数据库中的信息资源进行比较,确定所调查的四个树种所分出的48个形态类型,并鉴定46个类型到科和属或者到种的水平,其中与青海云杉共生的外生菌根真菌鉴定到科的水平的有2个类型,鉴定到属的有2个类型,其余7个类型都鉴定到了种;与白桦共生的外生菌根真菌鉴定到科的水平的有2个类型,鉴定到属的有4个类型,鉴定到种的有6个类型;与山杨共生的外生菌根真菌鉴定到科的有1个类型,鉴定到属的有3个类型,鉴定到种的有6个类型;与兴安落叶松共生的外生菌根真菌中鉴定到科的有3个类型,鉴定到属的有3个类型,鉴定到种的有7个类型;其中属于担子菌亚门的有36株,子囊菌亚门的有10株。本研究在进行分子鉴定的同时,充分的利用了互联网上的生物信息资源,构建了分类报告,并对其进行了分析。
综上所述,本研究中以宏观的形态、颜色和显微结构中菌套菌丝排列图式、菌套表面结构及菌丝和菌索内部结构等特征分出的48种类型,经DNA测序鉴定,基本上与测序得出结论相符,且结果表明,利用rDNA ITS区段进行分子测序,进而进行外生菌根真菌种类鉴定是切实可行的。
This research including morphological and anatomical characteristic analysis of different trees specises ectomycorrhizal types and study on molecular identification to these ectomycorrhizae. Now respectively abstract writes as follow:
1.Morphological and anatomical characters analysis of ectomycorrhizae
In this study, taking mycorrhizal sysbiosis as the study object, from one tree species starting and from the morphological and anatomical angle, studied these ectomycorrhizal types systematicly, and comprehensive characteristic of 48 ectomycorrhizal types were described using photographs and words. Among these types, there are 11 types on the Picea crassfolia, 13 types on the Betula platyphylla, 10 types on the Populus davidiana and 14 types on the Larix gmelinii. At the same times, Further made a primary identification on these types by using of ectomycorrhizal colour, morphological characters, surface structure of mantle and anatomical characters in plan views and so on; probed in the feasibility of ectomycorrhizal classification and identification in the light of exterior hyphae, mycorrhizal colour, interior microstructure such as arranged patterm of mantle hyphae, mantle surface structure and anatomical structure of hyphae and rhizomorphs mainly. On the basis, taxonomic key have been worked out on the different trees species.
2.Molecular identification reseach on the ectomycorrhizal root tips
This research was on the premise of having been sorted out basal ectomycorrhizal types on the basis of the morphological and anatomic ways, took the mycorrhizal symbiosis root tips of each tree species as studied material, extracted DNA of symbiosis, then ITS regions of DNA of identified ectomycorrhizal fungi were amplified using the special pairs of primers ITS1F /ITS4 and sequenced, the sequences determined were compared with those registered in rDNA sequence database on Internet, then identification results were obtained according to comparing results and other biological information in DNA sequence database on Internet. At last, ectomycorrhizal fungi diversity of several trees species in Inner mongolia were confirmed. The results write as follow: firstly, of the total of 48 morphology and anatomatical types of four trees species, 46 types were identified to the level of family, genus and species respectively, in addition, ectomycorrhizal types of Picea crassfolia belonging respectively to family level is 2, genus level is 2 and rest of 7 is identified to species level; ectomycorrhizal types of Betula platyphylla belonging respectively to family level is 2, genus level is 4, and species level is 6; ectomycorrhizal types of Populus davidiana belonging respectively to family level is 1,genus level is 3, and species level is 6; ectomycorrhizal types of Larix gmelinii belonging respectively to family level is 3, genus level is 3, and rest of 7 is identified to the species level. At the same times, among 46 identified ectomycorrhizal types, 36 species belong to Basidiomycotina and 10 species belong to Ascomycotina.
This research have finished species identifications, as well as by using the biological information resources on Internet adequately, constructed the taxonomy report, and analysed the compared results and taxonomy report.
In conclusion, 48 types that have been sorted out based on the macroscopical shape, colour and microcosmic structure, such as arranged patterm of mantle hyphae, mantle surface structure and anatomical structure of hyphae and rhizomorphs and so on was the same with the rusults using molecular identification. At the same times, it show that it is feasible using rDNA ITS regions to sequence and confirm these species.
(一)外生菌根形态学、解剖学特征分析
本研究从某个树种的角度出发,以菌根共生体为研究对象,系统地从形态学和解剖学的角度研究了与之共生的外生菌根类型,并以图谱及文字说明的形式对本研究实验中所采集到的菌根标本中的48种类型进行了详细描述;其中与青海云杉共生的外生菌根分出11种类型,与白桦共生的外生菌根分出13种类型,与山杨共生的外生菌根分出10种类型,与兴安落叶松共生的外生菌根14种类型。以宏观的形态、颜色和显微结构中菌套菌丝排列图式、菌套表面结构及菌丝和菌索内部结构等特征为主要依据进行外生菌根初步分类鉴定,并编制了检索表。
(二)外生菌根根尖分子鉴定研究
在依据形态学、解剖学特征分出基本类型的基础上,以各个树种的菌根共生体的根尖为材料,提取DNA,利用真菌特有引物ITS1F和ITS4,并在PCR基础上对外生菌根真菌rDNA的ITS区段进行碱基序列测定,然后与互联网上DNA序列数据库中的信息资源进行比较,确定所调查的四个树种所分出的48个形态类型,并鉴定46个类型到科和属或者到种的水平,其中与青海云杉共生的外生菌根真菌鉴定到科的水平的有2个类型,鉴定到属的有2个类型,其余7个类型都鉴定到了种;与白桦共生的外生菌根真菌鉴定到科的水平的有2个类型,鉴定到属的有4个类型,鉴定到种的有6个类型;与山杨共生的外生菌根真菌鉴定到科的有1个类型,鉴定到属的有3个类型,鉴定到种的有6个类型;与兴安落叶松共生的外生菌根真菌中鉴定到科的有3个类型,鉴定到属的有3个类型,鉴定到种的有7个类型;其中属于担子菌亚门的有36株,子囊菌亚门的有10株。本研究在进行分子鉴定的同时,充分的利用了互联网上的生物信息资源,构建了分类报告,并对其进行了分析。
综上所述,本研究中以宏观的形态、颜色和显微结构中菌套菌丝排列图式、菌套表面结构及菌丝和菌索内部结构等特征分出的48种类型,经DNA测序鉴定,基本上与测序得出结论相符,且结果表明,利用rDNA ITS区段进行分子测序,进而进行外生菌根真菌种类鉴定是切实可行的。
This research including morphological and anatomical characteristic analysis of different trees specises ectomycorrhizal types and study on molecular identification to these ectomycorrhizae. Now respectively abstract writes as follow:
1.Morphological and anatomical characters analysis of ectomycorrhizae
In this study, taking mycorrhizal sysbiosis as the study object, from one tree species starting and from the morphological and anatomical angle, studied these ectomycorrhizal types systematicly, and comprehensive characteristic of 48 ectomycorrhizal types were described using photographs and words. Among these types, there are 11 types on the Picea crassfolia, 13 types on the Betula platyphylla, 10 types on the Populus davidiana and 14 types on the Larix gmelinii. At the same times, Further made a primary identification on these types by using of ectomycorrhizal colour, morphological characters, surface structure of mantle and anatomical characters in plan views and so on; probed in the feasibility of ectomycorrhizal classification and identification in the light of exterior hyphae, mycorrhizal colour, interior microstructure such as arranged patterm of mantle hyphae, mantle surface structure and anatomical structure of hyphae and rhizomorphs mainly. On the basis, taxonomic key have been worked out on the different trees species.
2.Molecular identification reseach on the ectomycorrhizal root tips
This research was on the premise of having been sorted out basal ectomycorrhizal types on the basis of the morphological and anatomic ways, took the mycorrhizal symbiosis root tips of each tree species as studied material, extracted DNA of symbiosis, then ITS regions of DNA of identified ectomycorrhizal fungi were amplified using the special pairs of primers ITS1F /ITS4 and sequenced, the sequences determined were compared with those registered in rDNA sequence database on Internet, then identification results were obtained according to comparing results and other biological information in DNA sequence database on Internet. At last, ectomycorrhizal fungi diversity of several trees species in Inner mongolia were confirmed. The results write as follow: firstly, of the total of 48 morphology and anatomatical types of four trees species, 46 types were identified to the level of family, genus and species respectively, in addition, ectomycorrhizal types of Picea crassfolia belonging respectively to family level is 2, genus level is 2 and rest of 7 is identified to species level; ectomycorrhizal types of Betula platyphylla belonging respectively to family level is 2, genus level is 4, and species level is 6; ectomycorrhizal types of Populus davidiana belonging respectively to family level is 1,genus level is 3, and species level is 6; ectomycorrhizal types of Larix gmelinii belonging respectively to family level is 3, genus level is 3, and rest of 7 is identified to the species level. At the same times, among 46 identified ectomycorrhizal types, 36 species belong to Basidiomycotina and 10 species belong to Ascomycotina.
This research have finished species identifications, as well as by using the biological information resources on Internet adequately, constructed the taxonomy report, and analysed the compared results and taxonomy report.
In conclusion, 48 types that have been sorted out based on the macroscopical shape, colour and microcosmic structure, such as arranged patterm of mantle hyphae, mantle surface structure and anatomical structure of hyphae and rhizomorphs and so on was the same with the rusults using molecular identification. At the same times, it show that it is feasible using rDNA ITS regions to sequence and confirm these species.
引文
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