ArticlePDF Available

Effects of different combinations of carbon and nitrogen sources in MS medium on activities of extracellular β-1,3-glucanase produced by take-all pathogen

Authors:

Abstract and Figures

The pathogenic fungi Gaeumannomyces graminis var.tritici(Ggt) was cultured in modified MS medium(26 ℃,150 r/min) with different combinations of carbon and nitrogen sources.The enzyme activity in the culture filtrate was detected by 3,5-dinitrosalicylic acid(DNS) in 7,14,21 d after culturing the fungus.The results showed the enzyme activities were obviously different in the culture filtrate with wheat stalks,wheat leaves,maize stalks,maize leaves and maize roots as carbon source combined with different nitrogen sources.The highest enzyme activities were detected in the culture filtrate with wheat leaves as the carbon source and the lowest enzyme activities were in the culture filtrate with maize roots as the carbon source.The enzyme activities were not significantly different in the culture filtrate with wheat tissues as compared with that with maize tissues.Comparing 4 nitrogen sources,the enzyme activities were obviously higher in the culture filtrate with organic nitrogen than that with inorganic nitrogen.The highest enzyme activities were detected in the culture filtrate with beef extract as nitrogen source 7 d after culturing the fungus.The enzyme activities detected in the culture filtrate with peptone were low and reached the peak 21 d after culturing the fungus.The enzyme activities in the culture filtrate with beef extract decreased gradually during the culture stage.The best carbon and nitrogen combination in the medium for enzyme activities was beef exact and wheat leave detected 7 d after culturing the fungus.
Content may be subject to copyright.
35 卷 第 2西北农林科技大学学报(自然科学版) V ol. 35 N o. 2
2007 2Jou rn al of N or t hw est A & F U niver s ity ( N at . Sci. E d. ) Feb . 2007
不同碳、氮源组合对小麦全蚀病菌
产生胞外-1,3-葡聚糖酶的影响
余永廷,谢媛媛,黄丽丽,康振
(西北农林科技大学 植保学院,陕西 杨凌 712100)
  [摘 要] 利用改进的M S 液体培养基,通过改变碳、氮源组合 ,分别在振荡培养小麦全蚀病菌 7, 14, 2 1 d
( 26 , 150 r/min ) , 3, 5-二硝基水杨酸(DN S )比色法测定培养滤液中的
-1, 3-葡聚糖酶活性。结果表明,培养基
中分别以小麦茎秆、叶片,玉米茎秆、叶片和根系为碳源与不同氮源组合,培养滤液中-1, 3-葡聚糖酶活性有明显差
,其中以小麦叶片为碳源时酶活性最高,玉米根系为碳源时酶活性较低,但两者之间酶活性无明显差异。培养
基中加入有机氮时酶活性均明显高于加入无机氮,但两种有机氮培养基中的酶活性变化趋势完 全不同。以牛肉膏
为氮源时,在小麦全蚀病菌培养7d时培养滤液中的酶活性最高,且酶活性随病菌 培养时间的延长而逐渐下;
蛋白胨为氮源时酶活性则呈上升趋势,在病菌培养21d时最高。以上结果表明,供试各组合中以牛肉膏为氮源、
麦叶片为碳源的组合为最佳组合, 7 d时有相对较高的酶活性。
[关键词] 小麦全蚀病菌;禾顶囊壳小麦变种;-1, 3-葡聚糖酶; M S 培养基;碳源;氮源
[中图分类号] S4 35. 12 1     [文献标识码] A [ 文章编号] 1671-9 387 ( 2007) 02-01 10 -04
Effects of different combinations of carbon and nitrogen sources
in MS medium on activit ies of extracellular
-1, 3-
glucanase produced by take-
all pathogen
YU Y ong -t ing, X IE Yuan-yu an, HU AN G Li-li, K AN G Zhen-sheng
(College of Plant p r ote ct ion,N orth west A &F U niv ersi ty ,Y ang lin g,S ha anx i 712100, C hina)
Abstract: T h e p at hog enic f u ngi Gaeumannomy ces g rami nis var. trit ici ( Gg t ) w a s cult ur ed in m odif ied
M S mediu m ( 26 , 150 r/m in )wit h dif ferent combinat ion s of carb on and nit rog en sour ces.T he enzyme
activit y in t he cult u re f iltrat e w as det ected by 3, 5-
dinitros alicylic acid (DN S)in 7, 14, 21 d after cultu ring
the fungu s . T he result s show ed the enzy me activities w ere obviously d iff erent in t h e culture f ilt rat e w it h
w heat stalks, w he at leav es, m aize s t al ks, m aiz e lea ves and m aize r oot s as ca rbo n s ou rce com bin ed w it h
dif fere nt nit ro gen so urc es .T h e h ighest enzy me activit ies w ere detected in t he culture filt rat e w ith w heat
leaves as the carbon s ource and the low est enzyme activit ies w ere in t h e culture filtr ate w ith m aize roots as
the carbon s ource. T he enzyme activities w ere not significant ly different in t he cultu re filtrat e w ith w h eat
tissues as comp ared wit h th at w it h maize tissues. Com paring 4 nit ro gen sources, t he enzym e activities w ere
ob viou sly hig her in the cult u re f ilt rat e w it h o r gan ic n it ro g en t ha n t h at w it h inor g anic nit ro g en.T he h ighest
enzy me activit ies were detected in t he culture f ilt rate w ith beef ex t ract as nit rog en sou rce 7d aft er
cult u ring the fung u s. Th e enzym e act ivities detected in t he culture filt rat e wit h pept one were low and
[收稿日期] 2006-01-23
[基金项目] 国家自然科学基金项目( 30270863) ; 教育部“长江学者和创新团队”支持计划( PC S IRT 2 00558) ; 西北农林科技大学创新团队
资助项目
[作者简介] 余永廷( 1980- ) , ,安徽淮北人,在读硕士,主要从事小麦全蚀病致病机理研究。E -m ail: y uy o ngt ing@ 126. com
[通讯作者] 黄丽丽( 1961- ) , ,陕西周至人,教授,主要从事植物病害综合防治研究。E-mail: hua nglili1@ ho t mail. com
reached t h e peak 21 d after cult uring t h e fu ng us. T he enzy me activit ies in t he cult ure filtrat e w ith b eef
ext ract decreased gr aduall y during t he cult u re st ag e.T h e best ca rb on and n itr og en co mb in at io n in t he
me diu m fo r e nzym e act iv itie s w as be ef e x act and w heat leave det ect ed 7 d a fte r cu ltur ing t he f ung us .
Key words: t ake-all pat h ogen o f w heat ; Ga eum annomy ces gr aminis va r . tritici ;-1, 3-g lucanases; M S
medium;ca rbo n so urc e;nit rog en sour ce
  小麦全蚀病是由禾顶囊壳小麦变种
(G aeumannomyces g r amin is ( S acc. ) von . A rx an d
O livier v ar . tr itici Walk er) 侵染小麦引起的重要
茎部病害,主要引起黑根、分蘖减少、枯白等症状,
造成严重的产量和经济损失。由于该病在世界小麦
种植区广泛分布,且在小麦生育期均可危,之没
有免疫和高抗品种,使得病害防治更加困难。内外
植物病理工作者在小麦全蚀病的化学防治物防
致病机理等方面已经进行了大量的研究工
[ 1-8],基本明确了小麦全蚀病菌侵染过程的细胞
行为以及胞壁降解酶在致病过程中的作用[ 5-8] ,
别是对病菌分泌的纤维素酶、半纤维素酶果胶酶在
侵染、病过程中的作用进行了研究,但目前关于小
全蚀病菌分泌胞外
-1, 3-葡聚糖酶及其与致病过
程的关系研究报道甚少。
本课题组在对受小麦全蚀病菌侵染的根组织进
行细胞化学研究时证,小麦根组织受侵染后产生
的防御反应结构——“木质管(l ig n in -
tube)中含有
大量的
-1, 3-聚糖[ 9-10] ,此推测小麦全蚀病菌在
穿透木质管的过程中可能会分泌胞外-1, 3-聚糖
,以降解-1, 3-葡聚糖,达到穿透木质管而入侵、
扩展和定殖的目的。,采用液体发酵方法培养小
麦全蚀病菌,生物化学方法已经证实培养滤液中
含有病菌分泌的胞外-1, 3-葡聚糖酶[ 11] ,由于
酶产量少、活性低,分离纯化困难,所以目前仍未明
确其酶学性质、构及其功能。因此,本研究在前期
工作的基础上,选有利于产酶的培养基,一步
究小麦全蚀病菌胞-1, 3-葡聚糖酶的性质、
,揭示其在病菌侵染、致病过程中的作用奠定基
础。
1 材料与方法
1. 1 材 料
1. 1. 1  供试菌种 禾顶囊壳小麦变种
Gaeumannomy ce s gr aminis var .trit ici (Ggt ) 9826
,由西北农林科技大学植物病理研究所提供。
按常规方法在P DA 平板培养基上繁殖、保存备用。
1. 1. 2 基础培养基 液体发酵产酶培养基(改进的
M S 液体培养基) , V an Ho o f [ 12]
[ 11] 的方法。
1. 1. 3 供试碳源 1. 1. 2 改进的M S 液体培养基
的麸皮分别改为收获后晒干的小麦茎秆、叶片及
玉米根系、茎秆和叶片,量为1 %
1. 1. 4 供试氮源 1. 1. 2 改进的M S 液体培养基
的酵母浸膏分别改为牛肉膏白胨、硝酸铵
化铵,用量为1%
1. 1. 5 化学试剂及来源 昆布多糖( L am inar in)
S igm a 公司产品,其他试剂均为国产
1. 2 方 法
1. 2. 1 培养方法 将在PDA 平板培养基上黑暗培
( 26 ) 35 d 的小麦全蚀病菌,用打孔器取直径
5mm 的菌饼4,放入装有50 m L 液体培养基的
150 mL 三角瓶,3个重复,恒温摇床振荡培
( 26 ℃、150 r/ min, )并根据不同碳、源设
为不同处理。
1. 2. 2  酶液的提取 分别在振荡培养 7, 14, 21 d
,于无菌条件下吸取培养液, 4 10 000 r/m in
离心5 min, 取上清液备用。
1. 2. 3 酶活性的测定 酶活性的测定采3, 5-
硝基水杨酸(DNS)比色法。取0. 05 m o l/L pH 5. 5
的乙酸-乙酸缓冲液配制的 0. 1% 底物昆布多糖
100 L , 37 ℃水浴预热5 min 后加入10 0 L酶液,
混匀,37 保温30 min采用DNS 测定生成的
还原性糖[ 11]
一个酶活力单位(U)定义为: 37 ,min
底物昆布多糖1m ol 葡萄糖的酶量。
2 结果与分析
2. 1 以小麦组织为碳源对产酶的影响
  由图1可以看出,分别以小麦茎秆、叶片为碳源
与牛肉膏、蛋白胨、酸铵、氯化铵组合,培养滤液中
麦全蚀病菌胞外
-1, 3-葡聚糖酶活性在不同培
时间、不同组合之间明显不同。1a可以,
麦茎秆与牛肉膏组合,培养滤液中 -1, 3-葡聚糖
活性在病菌培养7 d 时较高,0. 062 U / mL ,
高于其他组, 14 d时酶活性略有下降( 0. 0 6 U/
111
2期 余永廷等:不同碳、氮源组合对小麦全蚀病菌产生胞外
-1, 3-葡聚糖酶的影响
mL ) , 但仍然高于其他组合,21d酶活性下降至
0. 051 U/mL ,较第7天下降了17. 7% 小麦茎秆
蛋白胨组,培养滤液中-1, 3-葡聚糖酶活性在
病菌培养过程中呈明显上升趋势, 7 d 时酶活性较
,0. 027 U/m L , 14 d时酶活性为0. 044 U/
m L , 较第 7天时增加了63. 0% , 21 d 时酶活性继
续上升,较第 7天时增加了 12 2. 2 % , 达到0. 06 U/
m L 。而以小麦茎秆与硝酸铵和氯化铵的组合,
滤液-1, 3-葡聚糖酶活性在整个病菌培养过程中
均较低,且酶活性变化不大。
1 以小麦组织为碳源时不同氮源对小麦全蚀病菌产生胞外-1, 3-葡聚糖酶的影响
-- . 牛肉膏; - - . 蛋白胨; - - . 硝酸铵; - - . 氯化铵;a.小麦茎秆;b.小麦叶片
Fig . 1 Ex tra ce llula r -1, 3-gluc an as e activit y det e cted in t he dif f ere nt m edia of wh eat s talk as the sin g le
C arbo n so u rce an d beef ex t ract , p ept one , N H 4
NO4and N H4Cl as ni tro g en so ur ce, respect ivel y , in t he diff e rent incu bat ion t ime
-- . Beef e x t ract ; - - . Pe pt o ne ; - - . amm onium nitr ate; - - . A mmonium chlo rine ; a. Wheat st alk; b . Whe at leaves
  由图 1 b 以看出,麦叶片与牛肉膏组,
滤液中-1, 3-葡聚糖酶活性在病菌培养过程中呈
逐渐下降趋势;小麦叶片与蛋白胨组合,滤液
-1, 3-葡聚糖酶活性在病菌培养过程中呈明显上
升趋势;以小麦叶片为碳源与牛肉膏蛋白胨组合
,养滤液中
-1, 3-葡聚糖酶活性在病菌培养过
程中均高于小麦茎秆与牛肉膏、蛋白胨组合。以小麦
叶片为碳源与硝酸铵和氯化铵组合,培养滤液中
-1, 3-葡聚糖酶活性在整个病菌培养阶段均较低,
酶活性变化不大。从图1还可以看出,以硝酸铵和
氯化铵为氮源时,-1 , 3 -葡聚糖酶活性在病菌培养
期间明显低于以牛肉膏和蛋白胨为氮源,说明有机
较无机氮有利于小麦全蚀病菌胞外
-1, 3-聚糖
酶的产生。
2. 2 以玉米组织为碳源对产酶的影响
由图2可以看出,以玉米茎秆、叶片及根系分别
为碳源,牛肉膏蛋白胨酸铵、氯化铵组合,
全蚀病菌胞外 -1, 3-葡聚糖酶活性在小麦全蚀病
菌不同培养时间、不同组合之间明显不同。从图2 a
可以看出,以玉米茎秆与牛肉膏组,培养滤液中-
1, 3-葡聚糖酶活性在病菌培养过程中呈逐渐下降趋
,7d时最高,0. 057 U/m L, 14 d后酶活性
明显下降, 21 d时酶活性下降至0. 03 U/mL ,较第7
天时下降了47. 4% , 且明显低于其他组;而以蛋白
为氮源的组合酶活性呈现升高趋势,7d时酶活
性为0. 037 U/mL , 1 4 d时达0. 049 U/mL ,较第7
时升高了13. 3% , 21 d酶活性上升至0. 075 U/
m L ,明显高于其他组合;硝酸铵及氯化铵为氮源
组合酶活性均较( 0. 0 210. 027 U/mL ) ,
菌培养过程中活性虽有上升,但幅度较小
玉米叶片为碳源与不同氮源组,在不同培
养时-1, 3-葡聚糖酶活性变化趋势以及酶活性水
平与以玉米茎秆为碳源时相似(2 b ) 以蛋白胨为
氮源, 21 d 以玉米叶片为碳源的酶活性比以玉米
秆为碳源的低10. 7% ; 以硝酸铵为氮源, 21 d
以玉米叶片为源的酶活性比以玉米茎秆为碳源的
11. 1 %
从图2c可以看出,玉米根系为碳源与牛肉膏
组合,
-1, 3-葡聚糖酶活性水平在不同时间与以玉
茎秆叶片为碳源的相,活性在14 d 时比7 d
时略高,之后明显下降;以蛋白胨为氮源的组合,
性水平在不同时间均较以玉米茎秆片为碳源
的组合低,21 d 时相差更明,分别比二者低35.
8% 42. 7% , 酶活性在培养期间增幅较小, 14
21 d 酶活性分别较7 d 时增长12. 9% 38. 7% ;
硝酸铵及氯化铵为氮源的组合,酶活性水平均比
以玉米茎秆、叶片为碳源的组合低,酶活性变化趋势
112 西北农林科技大学学报(自然科学版)35
也与二者不同。
2 以玉米组织为碳源时不同氮源对小麦全蚀病菌产生胞外
-1, 3-葡聚糖酶的影响
-- . 牛肉膏; - - . 蛋白胨; - - . 硝酸铵; - - . 氯化铵;a.玉米茎秆;b.玉米叶片;c.玉米根系
Fig . 2 Ext r a cellul ar -1, 3-g lucana se a ctiv it y d et ect ed in th e dif fe ren t media of ma ize s t alk as t he s ingle
C arbo n so u rce an d beef ex t ract , p ept one , N H 4
NO4and N H4Cl as ni tro g en so ur ce, respect ivel y , in t he diff e rent incu bat ion t ime
-- . Beef ex tr act; - - . P e pt o ne; - - . a mmo niu m n itr ate ; - - . A mmonium chlo rine;a.M aize st al k;b.M a ize l eav es ;c.M aize r oo t s
  从图2还可以看,以玉米茎秆、叶片及根系为
碳源,机氮明显较无机氮有利于小麦全蚀病菌胞
-1, 3-葡聚糖酶的产生;牛肉膏为氮,在病菌
培养早期酶活性较高;以蛋白胨为氮源,在病菌培养
后期酶活性较高;以硝酸铵和氯化铵为碳,论何
种供试氮源均不利于病菌产酶。
综合以上结果可以看出,小麦茎秆、叶片及玉
米茎秆、叶片和根系为碳源与不同氮源组合,-1, 3-
葡聚糖酶活性受碳源的影响较小而受氮源的影响较
,有机氮明显比无机氮有利。从培养时间来看,
牛肉膏为氮源时在培养初期即可获得较高的酶活
,较王旭丽等[ 11] 报道的培养时间缩2,更有
利于酶的分离纯化。
3 讨 论
小麦全蚀病菌侵染过程的细胞化学研究结果显
,病菌的侵染过程可能涉及-1, 3-葡聚糖酶[ 9-10] ,
病菌在液体培养过程中也能够产生胞外-1, 3-
聚糖[ 11] 。因此,为了分离纯化出足量的小麦全
菌胞
-1, 3-葡聚糖酶,明确该酶在病菌侵染过
程中的作用,及为抗病基因工程研究提供物质基
,首先应筛选出产酶培养基和利于小麦全蚀病菌
分泌胞外
-1, 3-葡聚糖酶的条件。
于不同植物病原真菌生长和产生胞外-1, 3-
葡聚糖酶的碳、源不同,如大麦葡聚糖或茯苓多糖
有利于里氏木(T rich od er ma ree se i)生胞外-
1, 3-葡聚糖酶,在以葡萄糖为碳的培养基中产酶
量较少[ 13] ;燕麦麸皮和酵母浸膏利于炭色旋孢腔
(Cochl iob ol us carbonum ),少量蔗糖则抑制其产
[ 12] ;San to s [ 14] 利用以葡萄糖和硝酸钠分别为
碳、氮源的查氏( Czapek) 培养基培养意大利青霉菌,
使其产生胞外-1, 3-葡聚糖酶,但同时研究发培养
基中过量的葡萄糖会制意大利青霉菌产酶,王旭丽
[ 11] 在研究小麦全蚀病菌产酶条件筛选时也发现,
葡萄糖和蔗糖抑制病菌分泌胞外-1, 3-葡聚,
以小麦麸皮和牛肉膏别为碳氮源的组合比他组
合更利于产酶。本研究发,试各组合中以牛肉膏
为氮源,小麦叶片为碳源的组合最好,在病菌培养早
-1, 3-葡聚糖酶活性就达到高水平这些研究结
果对前期的结果是很的补充和完善。
[参考文献]
[1] 陈怀古,王裕中,陈厚德,.小麦全蚀病药剂防治技术与应用
研究[J] . 江苏农业科学, 1997( 1) : 38-41.
[2] 王美南,商鸿生.华山新麦草对小麦全蚀病的抗病性研究 [ J] .
西北农林科技大学学报:自然科学版, 2000, 28( 6) : 49-71.
[3]  G arder P A , A ngus J F, Pitso n G D, e t a l. A com pa rison of six
mo th eds t o co ntr ol tak e-
all in w heat [J] . A us t ral ian Jour nal o f
Agr icu ltura l Research , 1998, 49( 8) : 1225-1240.
[4]  Co ok R J . P ro blem s and pr og res s i n t he biolo gical cont ro l o f
w heat t a ke-
all [J] . P la n Pat h, 1994, 43: 429-437.
[ 5 ]  D or i S , H ers henho rn J, So l el Z , e t a l. Ch ara cte rizatio n o f an
endopo l yg alac tu ro nase a ss ociate d w it h t ake-
all diseas e of
w heat [ J] . Physio l ogical and M ol ecula r Pla nt Pa t ho log y, 1992,
40( 3) : 203-210.
[6] Dori S , Solel Z, Ka shma n Y , e t a l. Ce ll w all-deg radin g e nzy mes
pro duced by G aeumanno myces gr aminis var. tr itici in v it ro and
in v iv o [J] . Phy siolog ical and M ol ecular P lant P at ho log y,
1995, 46( 3) : 189-198.
113
2期 余永廷等:不同碳、氮源组合对小麦全蚀病菌产生胞外
-1, 3-葡聚糖酶的影响
[7] Weste G .Ex t ra-
cellu lar enzyme pr oduction by various isolat es
of Op hio bolus gr aminis S acc [ J ] . Phy t opath Z, 1970a , 52: 204-
206.
[8] Weste G . E xt ra -ce llu lar enzyme pr oduction by vario us isolat es
of Op hio bolus g r am i nis and O.g ram ini s var.av enae .
En zy mes pro duced w ith in t he ho s t t is sue [J] . Ph y topa t h Z ,
1970b, 67: 327-336.
[9] Kang Z S , Hu ang L L, Buc henaue r H. Cyt o chemis tr y o f cell
w all com p on ent al tera ti ons in w h eat r oot s in fect ed by
G aeumnnnomyc es g rami nins var.trit ici [J] . J P l ant D is P r ot ec,
2000, 107( 4) : 337-351.
[ 10]  Huang L L , Kang Z S, Buc hen auer H. Co mpa r is on of infection
of w he at root s by P hial op hora gr aminicola and
Gaeum anno myces gr am inis var. tr it ici by ul trast ru ct ura l and
cyt och emical s tu dies [ J ] . J Plan Dis Prot e c, 2 001, 1 08 ( 6 ) :
593-607.
[11]  王旭丽,黄丽丽,康振生,.小麦全蚀病菌胞外 -1, 3-葡聚糖
酶的产生和部分特性的研究[ J ] . 菌物系统, 2003, 22( 4) : 628-
633.
[ 12]  V a n Ho of A ,Le y ka m J,Sc haeff er H J,et al.A sin g le
-1, 3-
g lucanase s ecreted by t he m aize pat hog en C ochlio bolus
c arbon um act s by a n e x oly tic m echa nism [ J ] . P hys io l M o lec
Pa t h, 1991, 39: 259-267.
[ 13]  Bamf ort h C W . T he adapt a bility, pur ificat ion, and p ropert ies
of exo-
-1, 3-
g lucana se fr om t he f ung us T r ic hoder ma
r eesei[J] . Bioc hem J , 1980, 191: 863-866.
[ 14]  Sa nt o s T , Vil la nue bv a J R, N om be la C. Pro du ction and
cat abolite r epre ssion o f P enic ill ium it ali cum bet a -g luc ana-
   s es[ J] . J B acterio l, 1977, 129( 1) : 52-58.
  (上接第 109 )
[参考文献]
[1] 张国海,郭香凤,史国安,.鲜食葡萄采后贮藏 研究进展[ J ] .
河南科技大学学报:农学版, 2003, 23( 3) : 31-34.
[2] 中国农业年鉴编辑委员.中国农业年鉴[M] . 北京:中国 农业
出版社, 2003.
[3] 潘春. 萄贮 [ J] . 葡萄培与 酿酒, 1992
( 3) : 32-34.
[4] 饶景萍,任小林.园艺产品贮运学[ M ] . 西安:陕西人民出版社,
2003: 226.
[ 5]  修 德任,周延 .红地球葡萄贮藏保鲜技术 [J] . 鲜与加 ,
2002( 5) : 24-26.
[6] 贺普超.葡萄学[ M ] . 北京:中国农业出版社, 1999: 217.
[7]  王如福,吴彩娥,范三红.采后GA32, 4-D 处理对葡萄贮藏效
果的影响[ J] . 山西农业大学学报, 2000, 20( 3) : 263-264.
[8] 欧阳寿如.葡萄的品种及其研究[M] . 太原:山西 人民出版社,
1980: 25.
[ 9]  薛应龙.植物生理 学实验手 [ M ] . 上海:上海科学技术出版
, 1985: 403-410.
[ 10]  于大永.无核白葡萄 浆果耐拉力、耐压力的研究[ D] . 陕西杨
:西北农林科技大学, 2003: 30-31.
[ 11]  吴德玲,修德任,张文恩,.葡萄浆果耐压力、果和果柄间
耐拉力与贮藏性关系的研究初报[ J ] . 萄 萄 科技, 1981,
17( 2) : 1-5.
[12]  刘春香,何启伟,于占东.黄瓜质地与组织结构、维素及果胶
含量的关系[J]中国蔬菜, 2003( 5) : 3-7.
114 西北农林科技大学学报(自然科学版)35
... The chitinase activity was determined by Tang [24]. The activity of β-1,3-glucanase was determined by the method of Yongting et al. [25]. ...
Article
Full-text available
ARTICLE INFO abstract Sugarcane smut disease, caused by Ustilago scitaminea Syd., significantly decreases yield in sugarcane production. Guangxi is the dominant sugarcane growing province in China, the leading sugarcane variety ROC22 suffers serious smut disease and results in considerable loss in the sugar industry. In this study, at the part of morphological structure, seedcane setts of sugarcane variety ROC22 was inoculated by dipping and grown in pots with soil, until it appeared smut whips, and then observed the appearance of sugarcane plants and the ultra-structure of bud cells in sugarcane. Settlings were inoculated by syringe and assessed ultra-structure and enzyme activities were assessed. The result indicated that infected sugarcane plant grew a long whip, and the cane stem was very tiny without economic value. The damage of smut infection on bud tissue is serious, the bud cells were out of shape, and the cell contents disappeared. The cell nucleus and nucleolocentrosome were deformated and disintegrated, the chromatin was not well-distributed on the cell nucleus and the nucleolocentrosome generated papillas. Most mitochondria were serious injured and disintegrated, and some of their ridges disappeared and became empty. Leaf samples were taken for the analyses of POD, SOD, PPO, chitinase and β-1,3-glucanase activity. The result showed that the activities of POD (+4.08%), SOD (+11.83%), PPO (+38.46%) and chitinase (+12.28%) were increased except β-1,3-glucanase (3.31%) in different degrees after the smut inoculation. The results suggest that after artificial needle inoculation, the increase percentage was found in different inoculation period.
... The chitinase activity was determined by Tang [24]. The activity of β-1,3-glucanase was determined by the method of Yongting et al. [25]. ...
... 0HDVXUHPHQW RI 62' 32' &+7 DQG */8 DFWLYLWLHV Leaves were collected after inoculation with RKNs at 0 h, 6 h, 12 h, 24 h, 72 h, 144 h, 10 d, and 15 d to measure the enzyme activities of SOD and POD (Zhao et al., 2000), CHT (Boller et al., 1983), and GLU (Yu et al., 2007). ...
Article
Full-text available
Root-knot nematodes (RKNs; Meloidogyne spp.) are obligate endoparasites that infect a large number of crop plants and cause severe yield losses. Tomato cultivars carrying the Mi-1 gene conferring root-knot nematode resistance have been widely used, but this gene loses its effectiveness at soil temperatures above 28°C. In this study, the mechanism of the loss of resistance to RKNs at high soil temperatures was examined using LA0655 (Solanum lycopersicum cv. Anahu), which contains the Mi-1 gene. It was found that high soil temperatures delayed the expression of the Mi-1 gene and reduced the activities of superoxide dismutase, peroxidase, chitinase, and β-1,3-glucanase. Although genes in the jasmonic acid (JA) pathway exhibited an obvious response at high soil temperatures, this response could not prevent the invasion of RKNs; indeed, at 30 days after inoculation with RKNs, the plants produced large numbers of root knots and egg masses at a soil temperature of 32°C. © 2015, Korean Society for Horticultural Science and Springer-Verlag GmbH.
... Leaves were collected after inoculation with RKNs at 0 h, 6 h, 12 h, 24 h, 72 h, 144 h, 10 d, and 15 d to measure the enzyme activities of SOD and POD (Zhao et al., 2000), CHT (Boller et al., 1983), and GLU (Yu et al., 2007). ...
Article
Full-text available
In recent years, root knot nematodes (RKN) are causing massive produce losses in vegetables. Grafting can effectively reduce the root knot nematode disease if proper stocks resistant to this disease are used. In this study, 11 tomato stocks from home and abroad were used to measure four kinds of protective enzymes including phenylalanine ammonia enzyme (PAL), polyphenol oxidase (PPO), peroxidase (POD) and superoxide dismutase (SOD), to differentiate the role of the protective enzymes play in the resistance to RKN. Results shows that 'LA0656' has the highest SOD level among all materials at 0.605 U/g FW, and broadly, the cultivars which have Mi gene have higher SOD activities and those without Mi gene have lower SOD activities. 'LA3601' have the highest levels of PPO activity, reaching 0.761 δ OD/(g•min) FW, followed by 'CH101' and 'LA0656', while 'LA2822' and 'LA2826' nearly get a 0. 'LA2822' have highest PAL levels among all the cultivars, reaching 5.444 U/g FW, and there are some cultivars which get extremely low PAL levels, such as 'LA0655', 'LA0656', 'LA3858' and 'HA4402'. POD enzyme levels of 'LA2822', 'LA3640' are almost 0, while the highest reached over 50δOD/(g•min) FW in 'LA0656' and 'LA2157'. Our results indicate a complex relationship between enzyme activities and resistance, to help enclose what role the protective enzymes play in resistance mechanism of different tomato cultivars, choose better stocks and it also provide theoretical basis for further breeding work.
T he adapt abilit y, pur ificat ion, and p ropert ies of exo--1, 3-g lucanase fr om t he f ung us T ric hod erma r eesei
  • C W Bamf Ort H
Bamf ort h C W. T he adapt abilit y, pur ificat ion, and p ropert ies of exo--1, 3-g lucanase fr om t he f ung us T ric hod erma r eesei[ J ]. Biochem J, 1980, 191: 863-866.
Comparis on of inf ection of w heat root s by P hial op hora gr am inicola and Gaeumannomyces gr aminis var. tr it ici by ul trast ru ct ural and cyt och emical s tu dies [ J ]
  • L L Huang
  • Z S Buchen Auer
Huang L L, K ang Z S, Buchen auer H. Comparis on of inf ection of w heat root s by P hial op hora gr am inicola and Gaeumannomyces gr aminis var. tr it ici by ul trast ru ct ural and cyt och emical s tu dies [ J ]. J Plan Dis Prot ec, 2001, 108 ( 6 ) : 593-607.
Ext ra-cellu lar enzyme product ion by various isolat es of Op hiobol us gr aminis S acc
  • G West E
West e G. Ext ra-cellu lar enzyme product ion by various isolat es of Op hiobol us gr aminis S acc[ J ]. Phyt opath Z, 1970a, 52: 204- 206.
Cyt ochemis tr y of cell w all comp on ent al terati on s in wh eat r oot s in fect ed by G aeumnnnomyc es g rami nins var. t rit ici [ J]
  • Z S Kang
  • L L Hu Ang
  • H Buchenauer
Kang Z S, Hu ang L L, Buchenauer H. Cyt ochemis tr y of cell w all comp on ent al terati on s in wh eat r oot s in fect ed by G aeumnnnomyc es g rami nins var. t rit ici [ J]. J Pl ant D is Prot ec, 2000, 107( 4) : 337-351.
A sin gle -1, 3- g lucanase s ecr et ed by t he maize pat hogen Coc hliobolus c arbonum act s by an exolyt ic mechanism
  • V An Hoof
  • A Leykam
  • J Schaef F Er
V an Hoof A, Leykam J, Schaef f er H J, et al. A sin gle -1, 3- g lucanase s ecr et ed by t he maize pat hogen Coc hliobolus c arbonum act s by an exolyt ic mechanism [ J ]. Phys iol M olec Pat h, 1991, 39: 259-267.
Produ ct ion and cat abolit e repression of P enic ill ium it ali cum bet a-glucanas es [ J]
  • T Sant Os
Sant os T, V il lanuebva J R, N ombela C. Produ ct ion and cat abolit e repression of P enic ill ium it ali cum bet a-glucanas es [ J]. J Bacteriol, 1977, 129( 1) : 52-58.