植物资源与环境学报
植物資源與環境學報
식물자원여배경학보
JOURNAL OF PLANT RESOURCES AND ENVIRONMENT
2014年
2期
27-33
,共7页
邓慧华%洪伟%吴承祯%谢安强%潘伟彬%段萍
鄧慧華%洪偉%吳承禎%謝安彊%潘偉彬%段萍
산혜화%홍위%오승정%사안강%반위빈%단평
马尾松%内生真菌%油脂含量%形态特征%ITS片段%菌种鉴定
馬尾鬆%內生真菌%油脂含量%形態特徵%ITS片段%菌種鑒定
마미송%내생진균%유지함량%형태특정%ITS편단%균충감정
Pinus massoniana Lamb.%endophytic fungus%lipid content%morphological characteristics%ITS fragment%fungus identification
采用组织块法从马尾松( Pinus massoniana Lamb.)根、茎和叶片中分离获得内生真菌,初步筛选出产油脂菌株且对其油脂含量进行了分析;并采用形态学及分子生物学方法对产油脂菌株进行了分类鉴定。结果显示:从马尾松植株中共获得21株内生真菌菌株,其中分离自根、茎和叶片的菌株分别为4株、6株和11株。有14株菌株的菌丝中有油滴;其中,分离自叶片的菌株ZP-1、分离自根的菌株ZP-2和分离自茎的菌株ZP-3的菌丝内油滴较多且油脂含量较高,平均油脂含量分别达到29.12%、25.03%和30.56%,差异极显著(P<0.01)。不同菌株的菌落颜色、菌丝和分生孢子形态特征明显不同,菌株ZP-1、ZP-2和ZP-3的形态分别与拟青霉属( Paecilomyces Bainier)、生赤壳菌属( Bionectria Speg.)和镰刀菌属( Fusarium Link)菌种的形态相似。与GenBank中相关真菌ITS序列的比对以及NJ系统树分析结果显示:菌株ZP-1与拟青霉属、菌株ZP-2与生赤壳菌属、菌株ZP-3与镰刀菌属间的ITS片段序列相似性均达到99%,在各自的NJ系统树上它们也分别聚在一起。初步确定菌株ZP-1、ZP-2和ZP-3分别属于拟青霉属、生赤壳菌属和镰刀菌属。
採用組織塊法從馬尾鬆( Pinus massoniana Lamb.)根、莖和葉片中分離穫得內生真菌,初步篩選齣產油脂菌株且對其油脂含量進行瞭分析;併採用形態學及分子生物學方法對產油脂菌株進行瞭分類鑒定。結果顯示:從馬尾鬆植株中共穫得21株內生真菌菌株,其中分離自根、莖和葉片的菌株分彆為4株、6株和11株。有14株菌株的菌絲中有油滴;其中,分離自葉片的菌株ZP-1、分離自根的菌株ZP-2和分離自莖的菌株ZP-3的菌絲內油滴較多且油脂含量較高,平均油脂含量分彆達到29.12%、25.03%和30.56%,差異極顯著(P<0.01)。不同菌株的菌落顏色、菌絲和分生孢子形態特徵明顯不同,菌株ZP-1、ZP-2和ZP-3的形態分彆與擬青黴屬( Paecilomyces Bainier)、生赤殼菌屬( Bionectria Speg.)和鐮刀菌屬( Fusarium Link)菌種的形態相似。與GenBank中相關真菌ITS序列的比對以及NJ繫統樹分析結果顯示:菌株ZP-1與擬青黴屬、菌株ZP-2與生赤殼菌屬、菌株ZP-3與鐮刀菌屬間的ITS片段序列相似性均達到99%,在各自的NJ繫統樹上它們也分彆聚在一起。初步確定菌株ZP-1、ZP-2和ZP-3分彆屬于擬青黴屬、生赤殼菌屬和鐮刀菌屬。
채용조직괴법종마미송( Pinus massoniana Lamb.)근、경화협편중분리획득내생진균,초보사선출산유지균주차대기유지함량진행료분석;병채용형태학급분자생물학방법대산유지균주진행료분류감정。결과현시:종마미송식주중공획득21주내생진균균주,기중분리자근、경화협편적균주분별위4주、6주화11주。유14주균주적균사중유유적;기중,분리자협편적균주ZP-1、분리자근적균주ZP-2화분리자경적균주ZP-3적균사내유적교다차유지함량교고,평균유지함량분별체도29.12%、25.03%화30.56%,차이겁현저(P<0.01)。불동균주적균락안색、균사화분생포자형태특정명현불동,균주ZP-1、ZP-2화ZP-3적형태분별여의청매속( Paecilomyces Bainier)、생적각균속( Bionectria Speg.)화렴도균속( Fusarium Link)균충적형태상사。여GenBank중상관진균ITS서렬적비대이급NJ계통수분석결과현시:균주ZP-1여의청매속、균주ZP-2여생적각균속、균주ZP-3여렴도균속간적ITS편단서렬상사성균체도99%,재각자적NJ계통수상타문야분별취재일기。초보학정균주ZP-1、ZP-2화ZP-3분별속우의청매속、생적각균속화렴도균속。
By means of tissue block method, endophytic fungi were obtained from root, stem and leaf of Pinus massoniana Lamb., and producing lipid strains were preliminarily selected out and their lipid contents were analyzed. And these lipid strains were also classified and identified by morphological and molecular biological methods. The results show that 21 strains of endophytic fungi have been obtained from P. massoniana, in which, the number of strains isolated from root, stem and leaf is 4, 6 and 11 strains, respectively. There are oil bodies in mycelia of 14 strains, in which, the number of oil body is more and lipid content is higher in mycelia of strain ZP-1 from leaf, strain ZP-2 from root and strain ZP-3 from stem, and average lipid content reaches to 29. 12%, 25. 03% and 30. 56%, respectively, with the extremely significant difference (P<0. 01). Colony color, morphological characteristics of mycelium and conidium of different strains are obviously different, and morphology of strains ZP-1, ZP-2 and ZP-3 is similar to that of fungus species in genera of Paecilomyces Bainier, Bionectria Speg. and Fusarium Link, respectively. The results of alignment with related fungus ITS sequences in GenBank and analysis of NJ phylogenetic tree show that the sequence similarity of ITS fragment of strain ZP-1 with Paecilomyces, strain ZP-2 with Bionectria, strain ZP-3 with Fusarium all reaches to 99%, and they are gathered respectively in their own NJ phylogenetic tree. It is preliminarily determined that strains ZP-1, ZP-2 and ZP-3 belong to genera of Paecilomyces, Bionectria and Fusarium, respectively.