Biodiversity Science ›› 2019, Vol. 27 ›› Issue (10): 1122-1131.doi: 10.17520/biods.2019180

• Original Papers • Previous Article     Next Article

Diversity of Cytospora chrysosperma from different hosts in Xinjiang

Zhao Ying1, Ma Rong1, Yin Yongxiang2, Zhang Zhidong3, Tian Chengming4, *()   

  1. 1 College of Forestry and Horticulture, Xinjiang Agricultural University, Urumqi 830052
    2 Forestry Research Institute, Xinjiang Agricultural University, Urumqi 830052
    3 Institute of Microbiology, Xinjiang Academy of Agricultural Sciences, Urumqi 830091
    4 Key Laboratory for Silviculture and Conservation of Ministry of Education College of Forestry, Beijing 100083
  • Received:2019-05-29 Accepted:2019-08-09 Online:2019-10-20
  • Tian Chengming E-mail:chengmt@bjfu.edu.cn

Cytospora chrysosperma is a very important pathogenic fungus that can be infects many different trees. Currently, the diversity, phylogenetic and genetic relationships of C. chrysosperma remain unclear as host differences could cause substantial genetic and diverse differences among C. chrysosperma strains. To analyze the genetic, phylogenetic, and current diversity of C. chrysosperma, physiological and biochemical characteristics and genetic tests were performed from different hosts in Xinjiang. Using molecular markings Biolog-FF and ISSR, 47 strains of C. chrysosperma have been divided into 15 types. Strains were cultured and displayed varying culture characteristics and carbon source utilization and metabolic capacity traits. Each strain tended to utilize more carbon over time, however, strains varied in their carbon source preferences and metabolic capacities. Using cluster analysis, 47 strains of C. chrysosperma divided into two clusters based on genetic similarity (0.58), suggesting genetic structure is essential to understanding C. chrysosperma diversity, as host species and geographical origins were not significant.

Key words: Cytospora chrysosperma, cultural characteristics, Biolog-FF, ISSR

Table 1

Related information of the isolates in this study"

菌株 Isolate 寄主 Host 采集时间 Collecting time 采集地点 Location
319-1 苹果 Malus sp. 2011/8/7 昌吉吉木萨尔县 Jimusar County, Changji Prefecture
790-2 苹果 Malus sp. 2015/6/28 塔城地区塔城市 Tacheng, Tacheng Prefecture
908 苹果 Malus sp. 2015/7/17 塔城地区乌苏市 Wusu, Tacheng Prefecture
936 苹果 Malus sp. 2015/8/4 塔城地区塔城市 Tacheng, Tacheng Prefecture
701 榆树 Ulmus sp. 2015/4/18 吐鲁番市鄯善县 Shanshan County, Turpan City
1155 榆树 Ulmus sp. 2015/8/20 昌吉州奇台县 Qitai County, Changji Prefecture
600-2 胡杨 Populus euphratica 2012/8/20 喀什地区泽普县 Zepu County, Kashgar Prefecture
626 新疆杨 Populus alba 2012/8/20 喀什地区喀什市 Kashgar City, Kashgar Prefecture
686-1 山杨 Populus davidiana 2014/8/31 塔城地区塔城市 Tacheng City, Tacheng Prefecture
686-2 山杨 Populus davidiana 2014/9/1 塔城地区塔城市 Tacheng City, Tacheng Prefecture
702 小叶杨 Populus simonii 2015/4/18 吐鲁番市鄯善县 Shanshan County, Turpan City
737 钻天杨 Populus nigra 2015/6/4 哈密地区哈密市 Hami City, Hami Prefecture
739 钻天杨 Populus nigra 2015/6/4 哈密地区哈密市 Hami City, Hami Prefecture
740 钻天杨 Populus nigra 2015/6/4 哈密地区哈密市 Hami City, Hami Prefecture
742 钻天杨 Populus nigra 2015/6/4 哈密地区哈密市 Hami City, Hami Prefecture
793 俄罗斯杨 Populus russkii 2015/6/29 塔城地区塔城市 Tacheng City, Tacheng Prefecture
882 北京杨 Populus beijingensis 2015/7/22 塔城地区乌苏市 Wusu City, Tacheng Prefecture
891-1 俄罗斯杨 Populus russkii 2015/7/17 塔城地区乌苏市 Wusu City, Tacheng Prefecture
891-3 俄罗斯杨 Populus russkii 2015/7/17 塔城地区乌苏市 Wusu City, Tacheng Prefecture
904 大叶杨 Populus lasiocarpa 2015/7/16 塔城地区乌苏市 Wusu City, Tacheng Prefecture
914-1 杨树 Populus sp. 2015/7/21 塔城地区乌苏市 Wusu City, Tacheng Prefecture
934 新疆杨 Populus alba 2015/8/3 塔城地区乌苏市 Wusu City, Tacheng Prefecture
1074-2 胡杨 Populus euphratica 2015/8/26 吐鲁番市托克逊县 Toxon County, Turpan City
1076 胡杨 Populus euphratica 2015/8/26 吐鲁番市托克逊县 Toxon County, Turpan City
812-1 柳树 Salix sp. 2015/7/1 塔城地区塔城市 Tacheng City, Tacheng Prefecture
835-1 柳树 Salix sp. 2015/7/11 塔城地区托里县 Tori County, Tacheng Prefecture
839-2 柳树 Salix sp. 2015/7/2 塔城地区塔城市 Tacheng City, Tacheng Prefecture
839-3 柳树 Salix sp. 2015/7/2 塔城地区塔城市 Tacheng City, Tacheng Prefecture
847 柳树 Salix sp. 2015/7/6 塔城地区裕民县 Yumin County, Tacheng Prefecture
856-1 疏锯齿柳 Salix serrulatifolia 2015/7/29 塔城地区沙湾县 Shawan County, Tacheng Prefecture
755-2 红瑞木 Swida alba 2015/6/4 哈密地区哈密市 Hami City, Hami Prefecture
874-2 旱柳 Salix matsudana 2015/7/26 塔城地区沙湾县 Shawan County, Tacheng Prefecture
895-2 柳树 Salix sp. 2015/7/17 塔城地区乌苏市 Wusu City, Tacheng Prefecture
896 灰柳 Salix cinerea 2015/7/14 塔城地区托里县 Tori County, Tacheng Prefecture
897 柳树 Salix sp. 2015/7/14 塔城地区托里县 Tori County, Tacheng Prefecture
901 疏锯齿柳 Salix serrulatifolia 2015/7/14 塔城地区托里县 Tori County, Tacheng Prefecture
919-1 旱柳 Salix matsudana 2015/7/22 塔城地区塔城市 Tacheng City, Tacheng Prefecture
933 金丝垂柳 Salix × aureo-pendula 2015/8/3 塔城地区塔城市 Tacheng City, Tacheng Prefecture
935 金丝垂柳 Salix × aureo-pendula 2015/8/3 塔城地区塔城市 Tacheng City, Tacheng Prefecture
942 旱柳 Salix matsudana 2015/8/4 塔城地区塔城市 Tacheng City, Tacheng Prefecture
944-2 柳树 Salix sp. 2015/8/10 哈密地区哈密市 Hami City, Hami Prefecture
976-1 柳树 Salix sp. 2015/8/10 五家渠市 Wujiaqu City
976-2 柳树 Salix sp. 2015/8/10 五家渠市 Wujiaqu City
863 旱柳 Salix matsudana 2015/7/29 塔城地区沙湾县 Shawan County, Tacheng Prefecture
921 柳树 Salix sp. 2015/7/31 塔城地区塔城市 Tacheng City, Tacheng Prefecture
922 疏锯齿柳 Salix serrulatifolia 2015/7/31 塔城地区塔城市 Tacheng City, Tacheng Prefecture
930-1 疏锯齿柳 Salix serrulatifolia 2015/8/3 塔城地区塔城市 Tacheng City, Tacheng Prefecture

Table 2

The sequence of ISSR primers used in this study"

编号
No.
引物
Primer
引物序列(5′-3′)
Primer sequence (5′-3′)
退火温度
Annealing temp. (℃)
1 IJ-5 GAGAGAGAGAGAGAGATG 52
2 IL-7 ACAACACACACACACAC 52
3 IJ-7 GGGTGGGGTGGGGTG 52
4 IJ-9 AGATGTGTGTGTGTGTG 52
5 IJ-8 ATAAGAGAGAGAGAGAG 52
6 ID-8 GTGCGTGCGTGCGTGC 52
7 IL-3 CACGAGAGAGAGAGAGA 52
8 IQ-2 CCAAACCACCACCACCACCA 53
9 IJ-3 AGAGAGAGAGAGAGAGTC 53
10 IJ-4 GAGAGAGAGAGAGAGATC 53

Table 3

Observation items and assignment of morphological characteristics of Cytospora chrysosperma on PDA in this study."

观察项 Item 代表值 Value
0 1 2 3 4 5
菌落颜色
Colony color
白色
White
米黄色
Light yellow
土黄色
Khaki
灰色
Gray
深灰色
Dark gray
桔红色
Orange
菌落形状
Colony shape
圆形生长
Circular growth
辐射状生长
Radial growth
- - - -
表面形态
Surface morphology
菌丝稠密
Hyphae dense
菌丝稠密
Hyphae dense
菌丝稠密
Hyphae dense
菌丝稠密
Hyphae dense
菌丝稠密
Hyphae dense
菌丝稀疏
Hyphae sparse
平伏
Flat exhibition
轻微凸起
Slight raised
轻微凸起
Slight raised
凸起
Raised
凸起
Raised
轻微凸起
Slight raised
无轮纹
No concentric ring
无轮纹
No concentric ring
有轮纹
Concentric ring
无轮纹
No concentric ring
有轮纹
Concentric ring
无轮纹
No concentric ring
有/无子实体
Sporulation
子实体小
Small sporophore
子实体小
Small sporophore
子实体大
Larger sporophore
子实体大
Larger sporophore
- -
密集型
Intensive
稀疏型
Sparse
稀疏型
Sparse
聚生型
Concentrated
- -
色素分泌
Pigment secretion
米白色
Creamy-white
米黄色
Light yellow
土黄色
Khaki
灰色
Gray
黑色
Black
桔红色
Orange

Fig. 1

The UPGMA dendrogram of 47 Cytospora chrysosperma isolates based on morphological characteristics. A-O, Branches with different culture traits. The numbers represent strain numbers, which correspond to those in Table 1."

Fig. 2

Changes in the amount of Cytospora chersosperma carbon source utilization and metabolic activity over time. The strain numbers are the same to Table 1."

Table 4

The utilization of carbon type by Cytospora chrysosperma in this study.The strain numbers are the same to Table 1."

碳源种类
Carbon source type
吸光度值 Absorbance value (Ci-R) 平均吸光
度值
Average absorbance value
812-1 897 1074-2 686-2 921 922 839-2 847 934 882 702 891-1 896 740 935
吐温80 Tween 80 0.735 0.715 0.296 0.525 0.334 0.472 0.332 0.305 0.349 0.290 0.598 0.458 0.401 0.812 0.700 0.488
苦杏仁苷 Amygdalin - 0.282 0.274 0.322 0.276 0.583 0.653 0.254 0.418 0.284 0.396 0.475 0.493 0.313 0.330 0.357
D-阿拉伯糖 D-Arabinose 0.685 0.632 0.821 0.521 0.266 0.709 0.624 0.250 0.419 0.272 0.499 0.557 0.425 0.426 0.307 0.494
L-阿拉伯糖 L-Arabinose 0.688 0.627 0.670 0.568 0.285 0.724 0.453 0.375 0.582 0.266 0.629 0.496 0.285 0.259 0.304 0.481
糊精 Dextrin 0.392 0.686 0.661 0.782 0.503 0.447 0.438 0.345 0.414 0.264 0.543 0.522 0.469 0.479 0.369 0.488
D-果糖 D-Fructose - 0.678 0.891 0.669 0.401 0.404 0.471 0.303 0.503 0.282 0.616 0.578 0.406 0.259 0.726 0.479
α-D-葡萄糖 α-D-glucose 0.385 0.669 0.500 0.766 0.334 0.379 0.572 0.301 0.347 0.464 0.618 0.689 0.535 0.287 0.551 0.493
D-甘露糖 D-Mannitol - 0.899 0.563 0.741 0.336 0.312 0.314 0.510 0.992 0.610 0.297 0.508 0.314 0.251 0.416 0.471
D-核糖 D-ribose 0.658 0.691 0.451 0.613 0.541 0.892 0.562 0.510 0.349 0.554 0.432 0.401 0.526 0.276 0.353 0.521
水杨苷 Salicin - 0.394 0.534 0.323 0.342 0.359 0.335 0.523 0.318 0.282 0.463 0.579 0.551 0.331 0.334 0.378
蔗糖 Sucrose - 0.28 0.430 0.255 0.349 0.498 - 0.305 0.349 - 0.356 0.252 - 0.343 0.518 0.262
D-木糖 D-xylose 0.756 0.686 0.270 0.592 0.336 0.518 0.423 0.347 0.319 0.666 0.293 0.592 0.406 0.356 0.514 0.472
富马酸 Fumaric acid - 0.354 0.361 0.378 0.436 0.381 - 1.522 0.314 0.595 0.280 0.625 - 0.294 0.517 0.404
琥珀酸 Butanedioic acid - 0.371 0.360 0.255 0.348 0.331 - 0.400 0.279 0.346 0.649 0.535 - 0.284 0.326 0.299
L-天门冬酰 L-asparagine - 1.246 0.460 0.578 0.436 0.304 0.306 0.453 0.258 0.329 0.284 0.410 0.260 0.265 0.954 0.436
平均吸光度值
Average absorbance value
0.287 0.614 0.503 0.526 0.368 0.488 0.366 0.447 0.414 0.367 0.464 0.512 0.338 0.349 0.481

Table 5

Strains and types of carbon sources that used carbon sources alone in this study"

菌株
Strain
碳水化合物
Carbohydrate
羧酸及其衍生物
Carboxylic acids and derivatives
氨基酸
Amino acid
磷酸盐类
Phosphates
胺类
Amine
1074-2 - - L-谷氨酸
L- glutamate
L-丙胺酰胺
L-alamide
-
847 - 溴代丁二酸
Bromosuccinic acid
- - -
934 - - - - 腺苷
Adenosin
891-1 - L-乳酸
L-lactic acid
L-苯丙氨酸
L- phenylalanine
- -
896 L-海藻糖
L-trehalose
- - - -
740 - ρ-羟基苯乙酸
ρ-hydroxy acid
琥珀酰胺酸
Succinamic acid
- -
L-天门冬氨酸
L-aspartic acid

Fig. 3

The UPGMA dendrogram of 47 Crtospora chrysosperma isolates with PCR-ISSR. The 47 strains were divided into group II and group II when genetic similarity coefficient was 0.58. Strain numbers see Table 1."

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