1 材料与方法
1.1 材料
1.2 方法
表1 扩增序列所用引物Tab. 1 Primers used for DNA sequences |
表2 序列登录号Tab. 2 Accession number of sequences in GeneBank |
| 种名 Species name | 菌株号 Strain No. | GeneBank登录号GenBank accession number | ||
|---|---|---|---|---|
| ITS | SSU | LSU | ||
| 实验菌株 | GSJF001 | MW193047.1 | MW227479.1 | MW227480.1 |
| Ganoderma australe | XJSJF003 | MN747808.1 | MN747807.1 | MN744680.1 |
| G. australe | YLJF001 | MN689631.1 | MN689632.1 | MN689633.1 |
| G. australe | MMHJF001 | MN696209.1 | MN696211.1 | MW131225.1 |
| G. sinense | GACP17101260 | MK313126.1 | MK341563.1 | MK336404.1 |
| G. sinense | SA17092559 | MK345452.1 | MK346845.1 | MK346834.1 |
| G. gibbosum | UB2 | KU569557.1 | KU695658.1 | KU570955.1 |
| G. gibbosum | SPC2 | KU569547.1 | KU695655.1 | KU570946.1 |
| G. nasalanense | LPDR17060211 | MK345441.1 | MK346842.1 | MK346831.1 |
| G. nasalanense | LPDR17060212 | MK345442.1 | MK346843.1 | MK346832.1 |
| G. lucidum | FK2580 | MT225392.1 | MT225549.1 | MT225545.1 |
| G. tropicum | Loe31 | MK007291.1 | MK235148.1 | MK215781.1 |
| G. orbiforme | LPDR14061420 | MK345447.1 | MK346844.1 | MK346833.1 |
| Phellinus nigricans | T.Niemela9065 | MF319077.1 | MF319008.1 | MF318934.1 |
1.3 数据处理
2 结果与分析
2.1 田间发病症状
图1 枸树茎腐病田间发病症状A：发病中期症状;B：发病后期症状;C：干旱季节病树茎干上生长的担子果;D：潮湿季节病树茎干上生长的担子果;E：病树茎基生长的担子果及表面的泥层状担孢子粉。 Fig. 1 Field symptoms of B. papyrifera stem rot A: The symptoms of middle period of disease; B: The symptoms of later period of disease; C: Basidiocarp that grow on the stems of diseased trees in the dry season; D: Basidiocarp that grow on the stems of diseased trees in the moist season; E: The basidiocarp and the muddy spore powder on the surface of the stems of diseased trees. |
2.2 致病性测定
图2 致病性测定及病菌担子果诱导A：接种幼树叶片失绿发黄;B：内部组织白腐;C、D：幼树接种病菌2个月后长出的担子果;E：对照;F：接种分离菌的瓶装木屑培养基诱导长出的担子果。 Fig. 2 Pathogenicity determination and basidocarpinduction A: Pathogens have colonized at the inoculation site; B: White rot appeared on the internal tissue; C and D: Basidocarps of Ganoderma that grows after young trees inoculated with pathogens after 2 months; E: CK; F: Isolated strains developed basidiocarps in bottled sawdust medium. |
2.3 病原菌鉴定
图3 病原菌形态A：菌落（正面）;B：潮湿条件下生长的担子果;C：干旱条件下生长的担子果（正面）D：担子果（反面）;E：担孢子;F、G：菌丝（①缠绕菌丝;②生殖菌丝;③骨架菌丝）。 Fig. 3 Morphology of pathogen A: Colony morphology (front side); B: Basidiocarps growing under moist conditions; C: Basidiocarp growing under dry conditions (front side); D: Basidiocarp (back side); E: Basidiospores; F and G: Hyphae (① Bindinghyphae; ② Generative hyphae; ③ Skeletal hyphae). |
图4 菌株GSJF001基于rDNA-ITS、LSU和SSU序列的系统发育树Fig. 4 Phylogenetic trees based on rDNA-ITS, LSUand SSU sequences of strains GSJF001 |
图5 菌株GSJF001在不同处理条件下的菌落直径不同小写字母表示差异显著（P<0.05）,不同大写字母表示差异极显著（P<0.01）。 Fig. 5 Colony diametet of GSJF001 under different treatment conditions Different lowercase letters represent significant difference (P<0.05), different uppercase letters represent extremely significant difference (P<0.01). |
2.4 不同温度、pH、光照条件对GSJF001菌株菌落生长的影响
2.5 不同培养基对GSJF001菌株菌落生长的影响
表3 不同培养基对GSJF001菌丝生长的影响Tab. 3 Effect of different culture media on GSJF001 mycelia growth |
| 培养基 Culture media | 菌落特征Colony characteristics | |
|---|---|---|
| 菌丝生长状态 Mycelium growth status | 菌落直径Colony diameter/cm | |
| PDA | 菌丝白色浓密,菌丝长,边缘整齐密集,呈放射状 | 7.27aA |
| CA | 菌丝白色较浓密,菌丝较长,边缘整齐密集,呈放射状 | 7.07bB |
| PSA | 菌丝白色浓密,菌丝长,边缘平展密集,呈放射状 | 6.80cC |
| CMA | 菌丝白色稀疏,菌丝较长,边缘整齐较密集,呈放射状 | 3.80eE |
| OMA | 菌丝半透明稀疏,菌丝较长,边缘平展稀疏,呈放射状 | 4.17dD |
| Richards | 菌丝半透明稀疏,菌丝长,边缘平展稀疏,呈放射状 | 2.67fF |
| 枸树茎干汁液 | 菌丝半透明稀疏,菌丝短,边缘整齐稀疏,呈放射状 | 4.07dD |
注：不同小写字母表示差异显著（P<0.05）,不同大写字母表示差异极显著（P<0.01）。 | |
Note: Different lowercase letters represent significant difference (P<0.05), different uppercase letters represent extremely significant difference (P<0.01). |