苹果炭疽菌低毒性菌株的筛选及控病效果的研究
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摘要
苹果炭疽菌(Colletotrichum gloeosporioides)对苹果采后造成严重经济损失,利用弱致病菌株先侵入寄主组织后诱发植物产生抗病性,可以减轻植物病害的发生和危害。通过离子注入和磁场照射的方法处理苹果炭疽菌,然后在苹果上进行活体筛选测定,获得低毒株,并对低毒菌株和强毒菌株的生物学特性进行比较,然后接种于苹果上,对其引起苹果体内的部分酶活的变化进行比较研究,主要研究结果如下:
1.低毒菌株的筛选和控病效果
1.1低毒菌株的筛选
本研究利用离子注入和磁场照射的方法处理苹果炭疽菌,通过调查菌株生长情况及接种在苹果上,对苹果上病斑的大小进行测定筛选,获得低毒株,发现离子注入C100-2-5低毒株和磁场处理C0.25-1-2低毒株对苹果有较好的保护作用。
1.2低毒性菌株对苹果炭疽病的控制效果
试验结果表明:通过离子注入获得的低毒性菌株对苹果的控制效果较好。在以不同比例混合接种的控病效果中,强毒性菌株与低毒性的菌株以1:2的比例时接种的效果较好。
2.低毒菌株和强毒菌株生物学特性的研究
研究了温度、pH值及碳、氮源对苹果炭疽病菌(Colletotrichum gloeosporioides)强毒性菌株及低毒性菌株的生长、产孢量和孢子萌发的影响,以及强毒菌株与低毒菌株致死温度的差异。
2.1温度对苹果炭疽病菌强毒性菌株及低毒性菌株的生长、产孢和孢子萌发的影响
结果表明:强毒菌株和低毒菌株的菌丝生长温度范围都是10～35℃,最适温度为28℃,但低毒菌株C100-2-5在温度较低的环境下生长速率较快,产生分生孢子的温度范围与分生孢子萌发的温度范围为15～35℃,低毒菌株的产孢量去低于强毒菌株的产孢量,但低毒菌株在低温下的孢子萌发率较高。
2.2 pH值对苹果炭疽病菌强毒性菌株及低毒性菌株的生长、产孢和孢子萌发的影响
低毒菌株和强毒菌株在pH 3～10的范围内均能生长和产孢,菌丝生长的最适pH值为4,产生分生孢子的最适pH值为4～5,但经离子注入的低毒性菌株C100-2-5的产孢量在不同的pH值下却一直低于经磁场处理的C0.25-1-2和强毒性菌株。
2.3碳源对苹果炭疽病菌强毒性菌株及低毒性菌株的生长、产孢和孢子萌发的影响
结果表明,在7种供试碳源中,各菌株在苹果果汁的培养基上菌丝生长最快,在甘露醇培养基上生长最慢;低毒菌株产孢量最多的是碳源为麦芽糖的培养基,而强毒菌株则在葡萄糖培养基上;以苹果果汁为碳源时菌丝生长良好,但不利于分生孢子的产生。在苹果果汁培养基上分生孢子的萌发率较高,低毒性菌株在D-果糖和甘露醇萌发率较低。
2.4氮源对苹果炭疽病菌强毒性菌株及低毒性菌株的生长、产孢和孢子萌发的影响
强毒菌株和低毒性菌株都在氮源为蛋白胨培养基上生长的最好,且孢子萌发率最高,与以其它为氮源培养基达到显著差异,但是在蛋白胨培养基上C100-2-5菌株没有C0.25-1-2菌株和强毒菌株生长的好,从产孢量来看,强毒菌株在尿素上产孢量最大,而两个低毒菌株都是在氮源为NH_NO_3的培养基上产孢量最大,在氮源为NH_4SO_4的培养基上产孢量最小。
2.5苹果炭疽病菌强毒性菌株及低毒性菌株致死温度
各菌株的分生孢子的致死温度为47℃15min或50℃5min。但菌丝的致死温度却不同,强毒菌株为60℃20min,C0.25-1-2为60℃25min,C100-2-5为60℃30min或65℃5min。
2.6苹果炭疽菌低毒性菌株遗传稳定性的测定
通过逐代培养低毒性苹果炭疽菌,然后分别接种到苹果果实上,活体测定病斑发生的大小,结果表明:各代之间的病斑直径没有达到差异显著,说明低毒性菌株具有相对的遗传稳定性。
3.苹果炭疽菌低毒性菌株致病机制研究
应用分光光度法对活体内外苹果炭疽菌的低毒菌株和强毒菌株产生的细胞壁降解酶进行活性分析,结果如下:
3.1活体外低毒菌株和强毒菌株产生的细胞壁降解酶活性的变化
研究结果表明,强毒菌株和低毒菌株之间变化趋势没有显著的差别,但各菌株之间分泌的PMG和C_X活性都达到差异显著。随着培养天数的增加苹果炭疽菌分泌的聚甲基半乳糖醛酸酶(PMG)的活性都在第5天和第8天有一个峰值,而羧甲基纤维素酶(C_X)活性先升后降,第5天达到峰值后即开始下降,到第9天又出现一次高峰后活性降低。
3.2活体内细胞壁降解酶活性的变化
苹果果实受侵染后,PMG和Cx活性都显著增高,强毒菌株和低毒菌株均先出现PMG活性高峰,后出现Cx活性高峰。强毒菌株在接种第5天PMG活性达到最高峰,而低毒菌株则在第4天达到最高峰。C100-2-5与C0.25-1-2的变化趋势相似。强毒菌株PMG活性总体上低于低毒菌株,而接种强毒菌株苹果果实cx活性总体上要高于接种低毒菌株的活性,且接种强毒菌株和低毒菌株的Cx活性都在第7天达到高峰。
4苹果对炭疽菌的抗性机制研究
4.1真菌细胞壁降解酶
苹果果实接种苹果炭疽菌的强毒菌株与低毒菌株后,苹果果实体内β-1,3-葡聚糖酶活性和几丁质酶活性都处于上升趋势,但整体上接种强毒菌株的酶活低于接种低毒菌株的酶活,未接菌的苹果果实体内的酶活性基本处于平稳状态,且活性很低。
4.2寄主主要防御酶
研究了采后苹果果实接种炭疽菌(Colletotrichum gloeosporioides)后,主要防御酶过氧化物酶(POD)、多酚氧化酶(PPO)和苯丙氨酸解氨酶(PAL)的活性变化。结果表明:三种酶的活性都在第4天达到最高峰,而后活性呈下降趋势。接种低毒菌株的苹果果实的POD酶活在前三天和第6天都高于接种于强毒菌株苹果以内的酶活,但在第4-5天却低于接种于强毒菌株苹果以内的酶活;而接种低毒菌株的苹果果实的PPO酶活一直低于接种于强毒菌株苹果以内的酶活。PAL活性的变化则是接种低毒菌株的酶活在前5天低于强毒菌株,第6天后则相反。不同的是菌株的毒性不同,引起果实体内酶的活性变化也略有不同。在接清水对照的苹果体内也存在这三种酶,但活性很低。
4.3糖类
通过测定接种前后果实中可溶性总糖的含量,分析了可溶性总糖的变化。研究表明,接种强毒菌株苹果果实糖含量整体上低于接种低毒菌株的含量,都比未接种的清水对照糖含量高。
Apple anthracnose caused by Colletotrichum gloeosporioides has been an important disease of postharvest apple, which caused fruit rot during storage. In this paper, after used the hypo virulence strains invade the host organization to induce the plant to produce disease-resistant, may reduce the occurrence of the plant disease. Screening hypo virulence strains through ion implantation and the magnetic field, carries on the determination to it on the apple protective function, obtained hypo virulence, and the comparison to the strain biology characteristic between the the hypo virulence strains and virulence strain, then comparison the enzymes live change after inoculate on the apple. The experimental results were summarized as follows:
1. Screening hypo virulence strains and control effect
1.1. Screening hypo virulence strains
Screening hypo virulence strains through ion implantation and the magnetic field ,and research the mycelium strain growth situation and sickness spotsize which occurs on the apple, obtains hypo virulence, discovered ion implantation C100-2-5 is hypo virulence and the magnetic field processes C0.25-1-2 hypo virulence to have the better protective function to the apple.
1.2 The control effect of hypo virulence strain inoculated to apple
The test result indicated that, The hypo virulence strain obtains which through ion implantation the control effect is better to the apple. In controls effect by the different proportion simultaneous inoculation, the effect virulence strain and hypo virulence strain by 1: 2 proportions vaccinates is better.
2. Research on biology characteristic of hypo virulence strains and virulence strain
Had studied the influence of temperature, the PH value and the carbon, N sources to virulence the strain and the hypo virulence strain growth, the spore ,the spore germinstion and sporulation.
2.1 Effects of temperature on mycelial growth, spore germination and sporulation of Colletotrichum gloeosporioides
The result indicated that, the mycelium growth temperature range from10 to 35℃, the optimum temperature is 28℃, but hypo virulence strain C100-2-5 growth speed quicker under a lower temperature, the temperature of product the spore and spore germination is from 15 to 35℃, hypo virulence strain spore sprouting rate is higher at lower temperature.
2.2 Effects of pH on mycelial growth, spore germination and sporulation of Colletotrichum gloeosporioides
The mycelium growth most suitable pH value basic is same, can grow in the pH 3～10 scope and sporulation, the most suitable pH value is 4, and spore sprouting is 4-5, but produces the spore quantity after ion implantation hypo virulence strain C100-2-5 ,actually continuously is lower than under the different pH value and the temperature after magnetic field processing C0.25-1-2 and the virulence strain.
2.3 Effects of carbon sources on mycelial growth, spore germination,sporulation of Colletotrichum gloeosporioides.
The result indicated that, in 7 kinds carbon sources center, the mycelium growth quickest is in apple fruit juice, growth on the mannite culture medium is slowly, but hypo virulence strains produces the spore quantity many is the maltose, virulence strains produces the spore quantity in the glucose. Take apple fruit juice as carbon sources when the mycelium growth is good, But is disadvantageous to the spore production.
On the apple fruit juice culture medium spore sprouting rate is higher,the hypo virulence strain in the D- fructose and mannite sprouting rate is lower.
2.4 Effects of N sources on mycelial growth, spore germination、sporulation of Colletotrichum gloeosporioides
The virulence strain and the hypo virulence strain all in the proteinpeptone culture medium grow well, also spore sprouting rate is highest, achieves the remarkable difference with other culture medium, but theC100-2-5 strain does not have good on the protein peptone culture medium which the CO.25-1-2 strain and virulence strain grows, the virulence strain produces the sporequantity on the urea in a big way, both two hypo virulence strains are produce the spore quantity on NH4NO3 to be biggest,on the NH4SO4 is the smallest
2.5 Virulence strain and hypo virulence strain lethal temperature
The spore lethal temperature is 47℃15min or 50℃5min. But mycelium the lethal temperature is actually different, the virulence strain is60℃20min, C0.25-1-2 is 60℃25min, C100-2-5 is 60℃30min or 65℃5min.
2.6 Heredity stability of Hypo virulence strains
Through chases the generation to raise the hypo virulence strain,then separately vaccinates on the apple fruit, determination sickness spot occurs the size, the result indicated: Each generation of between sickness spot diameter had not achieved the difference is remarkable, showed the hypo virulence strain has the relative hereditystability.
3. Study on the Pathogenesis of Virulence strain and hypo virulence strain
The application spectrophoto metric method the cell wall which the hypo virulence strains and the virulence strain produces to the living specimen inside and outside degrades the enzyme to carry on the active analysis,the result as follow:
3.1 Change in the enzyme activity produces the cell wall degrades living specimens betweenthe hypo virulence strains and the virulence strain
The findings indicated that, between the virulence strain and the hypo virulence strain not the remarkable difference, but between various strains secretes PMG and the CX activeness all achieved the difference is remarkable. gathers the methyl galacturonic acid enzyme along with the raise number of days increaseapple anthrax secretion (PMG) activeness all has a peak value in 5thdays, after the carboxy methylcellulose enzyme (CX) activeness firstrises to fall, after 5th days achieved the peak value namely starts to drop, appears a time of peak after ninth days activeness to reduce.
3.2 Change in living specimens the cell wall degrades the enzyme activity
After the apple fruit is invaded dyes, the PMG and the Cx all activeness remarkably advances, the virulence strain and the hypo virulence strain first appear the PMG active peak, latter appears the Cx active peak. the virulence strain is vaccinating the 5th day enzyme exactly to achieve the high point, while the hypo virulence strain at 4th day. C100-2-5 and the C0.25-1-2 change tendency is similar.the virulence strain in the strain PMG active overall is lower than the hypo virulence strain, but vaccinates the virulence strain Cx active is higher than vaccinates the hypo virulence strain, also the vaccination the virulence strain and the hypo virulence strain Cx activeness all achieves the peak in 7th days.
4 Research on main enzyme
After the apple fruit inoculate the virulence strain and the hypo virulence strains, the apple fruit inβ-1,3- glucosan enzyme and several Ding nature enzyme activity all is in the trend of escalation, inoculate the virulence strain enzyme exactly is lower than the hypo virulence strain the CK in vivo enzyme activity basically to be at the steady condition, also activeness is very low
After studied the POD, PPO and PAL active change. The result indicated that, Three kind of enzymes activeness all achieve the high point in 4th days, but after activeness assumes the drop tendency. Vaccinates the hypo virulence strain on apple fruit, before three days and at 6th days all POD enzyme is higher than the virulence strain, but actually is lower than the virulence strain atthe 4-5th day; But vaccinates the hypo virulence strain apple fruit PPO enzyme to live continuously is lower than the virulence strain. The PAL active change is vaccinates the the hypo virulence strain enzyme to live in previous 5 days is lower than the virulence strain, after 6th days then is opposite. Different is the strain toxicity is different,causes the fruit in vivo enzyme active change also slightly to have the difference. In comparison apple also have these three kind of enzymes, but activeness is very low.
Through the determination sugar content, has analyzed the soluble total sugar change. The search indicated that, in the vaccination the virulence strain strain apple fruit sugar content whole is lower than vaccinates the the hypo virulence strain content, but all higher than comparison.
1.低毒菌株的筛选和控病效果
1.1低毒菌株的筛选
本研究利用离子注入和磁场照射的方法处理苹果炭疽菌,通过调查菌株生长情况及接种在苹果上,对苹果上病斑的大小进行测定筛选,获得低毒株,发现离子注入C100-2-5低毒株和磁场处理C0.25-1-2低毒株对苹果有较好的保护作用。
1.2低毒性菌株对苹果炭疽病的控制效果
试验结果表明:通过离子注入获得的低毒性菌株对苹果的控制效果较好。在以不同比例混合接种的控病效果中,强毒性菌株与低毒性的菌株以1:2的比例时接种的效果较好。
2.低毒菌株和强毒菌株生物学特性的研究
研究了温度、pH值及碳、氮源对苹果炭疽病菌(Colletotrichum gloeosporioides)强毒性菌株及低毒性菌株的生长、产孢量和孢子萌发的影响,以及强毒菌株与低毒菌株致死温度的差异。
2.1温度对苹果炭疽病菌强毒性菌株及低毒性菌株的生长、产孢和孢子萌发的影响
结果表明:强毒菌株和低毒菌株的菌丝生长温度范围都是10～35℃,最适温度为28℃,但低毒菌株C100-2-5在温度较低的环境下生长速率较快,产生分生孢子的温度范围与分生孢子萌发的温度范围为15～35℃,低毒菌株的产孢量去低于强毒菌株的产孢量,但低毒菌株在低温下的孢子萌发率较高。
2.2 pH值对苹果炭疽病菌强毒性菌株及低毒性菌株的生长、产孢和孢子萌发的影响
低毒菌株和强毒菌株在pH 3～10的范围内均能生长和产孢,菌丝生长的最适pH值为4,产生分生孢子的最适pH值为4～5,但经离子注入的低毒性菌株C100-2-5的产孢量在不同的pH值下却一直低于经磁场处理的C0.25-1-2和强毒性菌株。
2.3碳源对苹果炭疽病菌强毒性菌株及低毒性菌株的生长、产孢和孢子萌发的影响
结果表明,在7种供试碳源中,各菌株在苹果果汁的培养基上菌丝生长最快,在甘露醇培养基上生长最慢;低毒菌株产孢量最多的是碳源为麦芽糖的培养基,而强毒菌株则在葡萄糖培养基上;以苹果果汁为碳源时菌丝生长良好,但不利于分生孢子的产生。在苹果果汁培养基上分生孢子的萌发率较高,低毒性菌株在D-果糖和甘露醇萌发率较低。
2.4氮源对苹果炭疽病菌强毒性菌株及低毒性菌株的生长、产孢和孢子萌发的影响
强毒菌株和低毒性菌株都在氮源为蛋白胨培养基上生长的最好,且孢子萌发率最高,与以其它为氮源培养基达到显著差异,但是在蛋白胨培养基上C100-2-5菌株没有C0.25-1-2菌株和强毒菌株生长的好,从产孢量来看,强毒菌株在尿素上产孢量最大,而两个低毒菌株都是在氮源为NH_NO_3的培养基上产孢量最大,在氮源为NH_4SO_4的培养基上产孢量最小。
2.5苹果炭疽病菌强毒性菌株及低毒性菌株致死温度
各菌株的分生孢子的致死温度为47℃15min或50℃5min。但菌丝的致死温度却不同,强毒菌株为60℃20min,C0.25-1-2为60℃25min,C100-2-5为60℃30min或65℃5min。
2.6苹果炭疽菌低毒性菌株遗传稳定性的测定
通过逐代培养低毒性苹果炭疽菌,然后分别接种到苹果果实上,活体测定病斑发生的大小,结果表明:各代之间的病斑直径没有达到差异显著,说明低毒性菌株具有相对的遗传稳定性。
3.苹果炭疽菌低毒性菌株致病机制研究
应用分光光度法对活体内外苹果炭疽菌的低毒菌株和强毒菌株产生的细胞壁降解酶进行活性分析,结果如下:
3.1活体外低毒菌株和强毒菌株产生的细胞壁降解酶活性的变化
研究结果表明,强毒菌株和低毒菌株之间变化趋势没有显著的差别,但各菌株之间分泌的PMG和C_X活性都达到差异显著。随着培养天数的增加苹果炭疽菌分泌的聚甲基半乳糖醛酸酶(PMG)的活性都在第5天和第8天有一个峰值,而羧甲基纤维素酶(C_X)活性先升后降,第5天达到峰值后即开始下降,到第9天又出现一次高峰后活性降低。
3.2活体内细胞壁降解酶活性的变化
苹果果实受侵染后,PMG和Cx活性都显著增高,强毒菌株和低毒菌株均先出现PMG活性高峰,后出现Cx活性高峰。强毒菌株在接种第5天PMG活性达到最高峰,而低毒菌株则在第4天达到最高峰。C100-2-5与C0.25-1-2的变化趋势相似。强毒菌株PMG活性总体上低于低毒菌株,而接种强毒菌株苹果果实cx活性总体上要高于接种低毒菌株的活性,且接种强毒菌株和低毒菌株的Cx活性都在第7天达到高峰。
4苹果对炭疽菌的抗性机制研究
4.1真菌细胞壁降解酶
苹果果实接种苹果炭疽菌的强毒菌株与低毒菌株后,苹果果实体内β-1,3-葡聚糖酶活性和几丁质酶活性都处于上升趋势,但整体上接种强毒菌株的酶活低于接种低毒菌株的酶活,未接菌的苹果果实体内的酶活性基本处于平稳状态,且活性很低。
4.2寄主主要防御酶
研究了采后苹果果实接种炭疽菌(Colletotrichum gloeosporioides)后,主要防御酶过氧化物酶(POD)、多酚氧化酶(PPO)和苯丙氨酸解氨酶(PAL)的活性变化。结果表明:三种酶的活性都在第4天达到最高峰,而后活性呈下降趋势。接种低毒菌株的苹果果实的POD酶活在前三天和第6天都高于接种于强毒菌株苹果以内的酶活,但在第4-5天却低于接种于强毒菌株苹果以内的酶活;而接种低毒菌株的苹果果实的PPO酶活一直低于接种于强毒菌株苹果以内的酶活。PAL活性的变化则是接种低毒菌株的酶活在前5天低于强毒菌株,第6天后则相反。不同的是菌株的毒性不同,引起果实体内酶的活性变化也略有不同。在接清水对照的苹果体内也存在这三种酶,但活性很低。
4.3糖类
通过测定接种前后果实中可溶性总糖的含量,分析了可溶性总糖的变化。研究表明,接种强毒菌株苹果果实糖含量整体上低于接种低毒菌株的含量,都比未接种的清水对照糖含量高。
Apple anthracnose caused by Colletotrichum gloeosporioides has been an important disease of postharvest apple, which caused fruit rot during storage. In this paper, after used the hypo virulence strains invade the host organization to induce the plant to produce disease-resistant, may reduce the occurrence of the plant disease. Screening hypo virulence strains through ion implantation and the magnetic field, carries on the determination to it on the apple protective function, obtained hypo virulence, and the comparison to the strain biology characteristic between the the hypo virulence strains and virulence strain, then comparison the enzymes live change after inoculate on the apple. The experimental results were summarized as follows:
1. Screening hypo virulence strains and control effect
1.1. Screening hypo virulence strains
Screening hypo virulence strains through ion implantation and the magnetic field ,and research the mycelium strain growth situation and sickness spotsize which occurs on the apple, obtains hypo virulence, discovered ion implantation C100-2-5 is hypo virulence and the magnetic field processes C0.25-1-2 hypo virulence to have the better protective function to the apple.
1.2 The control effect of hypo virulence strain inoculated to apple
The test result indicated that, The hypo virulence strain obtains which through ion implantation the control effect is better to the apple. In controls effect by the different proportion simultaneous inoculation, the effect virulence strain and hypo virulence strain by 1: 2 proportions vaccinates is better.
2. Research on biology characteristic of hypo virulence strains and virulence strain
Had studied the influence of temperature, the PH value and the carbon, N sources to virulence the strain and the hypo virulence strain growth, the spore ,the spore germinstion and sporulation.
2.1 Effects of temperature on mycelial growth, spore germination and sporulation of Colletotrichum gloeosporioides
The result indicated that, the mycelium growth temperature range from10 to 35℃, the optimum temperature is 28℃, but hypo virulence strain C100-2-5 growth speed quicker under a lower temperature, the temperature of product the spore and spore germination is from 15 to 35℃, hypo virulence strain spore sprouting rate is higher at lower temperature.
2.2 Effects of pH on mycelial growth, spore germination and sporulation of Colletotrichum gloeosporioides
The mycelium growth most suitable pH value basic is same, can grow in the pH 3～10 scope and sporulation, the most suitable pH value is 4, and spore sprouting is 4-5, but produces the spore quantity after ion implantation hypo virulence strain C100-2-5 ,actually continuously is lower than under the different pH value and the temperature after magnetic field processing C0.25-1-2 and the virulence strain.
2.3 Effects of carbon sources on mycelial growth, spore germination,sporulation of Colletotrichum gloeosporioides.
The result indicated that, in 7 kinds carbon sources center, the mycelium growth quickest is in apple fruit juice, growth on the mannite culture medium is slowly, but hypo virulence strains produces the spore quantity many is the maltose, virulence strains produces the spore quantity in the glucose. Take apple fruit juice as carbon sources when the mycelium growth is good, But is disadvantageous to the spore production.
On the apple fruit juice culture medium spore sprouting rate is higher,the hypo virulence strain in the D- fructose and mannite sprouting rate is lower.
2.4 Effects of N sources on mycelial growth, spore germination、sporulation of Colletotrichum gloeosporioides
The virulence strain and the hypo virulence strain all in the proteinpeptone culture medium grow well, also spore sprouting rate is highest, achieves the remarkable difference with other culture medium, but theC100-2-5 strain does not have good on the protein peptone culture medium which the CO.25-1-2 strain and virulence strain grows, the virulence strain produces the sporequantity on the urea in a big way, both two hypo virulence strains are produce the spore quantity on NH4NO3 to be biggest,on the NH4SO4 is the smallest
2.5 Virulence strain and hypo virulence strain lethal temperature
The spore lethal temperature is 47℃15min or 50℃5min. But mycelium the lethal temperature is actually different, the virulence strain is60℃20min, C0.25-1-2 is 60℃25min, C100-2-5 is 60℃30min or 65℃5min.
2.6 Heredity stability of Hypo virulence strains
Through chases the generation to raise the hypo virulence strain,then separately vaccinates on the apple fruit, determination sickness spot occurs the size, the result indicated: Each generation of between sickness spot diameter had not achieved the difference is remarkable, showed the hypo virulence strain has the relative hereditystability.
3. Study on the Pathogenesis of Virulence strain and hypo virulence strain
The application spectrophoto metric method the cell wall which the hypo virulence strains and the virulence strain produces to the living specimen inside and outside degrades the enzyme to carry on the active analysis,the result as follow:
3.1 Change in the enzyme activity produces the cell wall degrades living specimens betweenthe hypo virulence strains and the virulence strain
The findings indicated that, between the virulence strain and the hypo virulence strain not the remarkable difference, but between various strains secretes PMG and the CX activeness all achieved the difference is remarkable. gathers the methyl galacturonic acid enzyme along with the raise number of days increaseapple anthrax secretion (PMG) activeness all has a peak value in 5thdays, after the carboxy methylcellulose enzyme (CX) activeness firstrises to fall, after 5th days achieved the peak value namely starts to drop, appears a time of peak after ninth days activeness to reduce.
3.2 Change in living specimens the cell wall degrades the enzyme activity
After the apple fruit is invaded dyes, the PMG and the Cx all activeness remarkably advances, the virulence strain and the hypo virulence strain first appear the PMG active peak, latter appears the Cx active peak. the virulence strain is vaccinating the 5th day enzyme exactly to achieve the high point, while the hypo virulence strain at 4th day. C100-2-5 and the C0.25-1-2 change tendency is similar.the virulence strain in the strain PMG active overall is lower than the hypo virulence strain, but vaccinates the virulence strain Cx active is higher than vaccinates the hypo virulence strain, also the vaccination the virulence strain and the hypo virulence strain Cx activeness all achieves the peak in 7th days.
4 Research on main enzyme
After the apple fruit inoculate the virulence strain and the hypo virulence strains, the apple fruit inβ-1,3- glucosan enzyme and several Ding nature enzyme activity all is in the trend of escalation, inoculate the virulence strain enzyme exactly is lower than the hypo virulence strain the CK in vivo enzyme activity basically to be at the steady condition, also activeness is very low
After studied the POD, PPO and PAL active change. The result indicated that, Three kind of enzymes activeness all achieve the high point in 4th days, but after activeness assumes the drop tendency. Vaccinates the hypo virulence strain on apple fruit, before three days and at 6th days all POD enzyme is higher than the virulence strain, but actually is lower than the virulence strain atthe 4-5th day; But vaccinates the hypo virulence strain apple fruit PPO enzyme to live continuously is lower than the virulence strain. The PAL active change is vaccinates the the hypo virulence strain enzyme to live in previous 5 days is lower than the virulence strain, after 6th days then is opposite. Different is the strain toxicity is different,causes the fruit in vivo enzyme active change also slightly to have the difference. In comparison apple also have these three kind of enzymes, but activeness is very low.
Through the determination sugar content, has analyzed the soluble total sugar change. The search indicated that, in the vaccination the virulence strain strain apple fruit sugar content whole is lower than vaccinates the the hypo virulence strain content, but all higher than comparison.
引文
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