丽格海棠抗茎腐病的生物学研究及EMS诱导的突变体筛选
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摘要
丽格海棠,学名Begonia×hiemalis,英文名Rieger begonia,别名玫瑰海棠,属于秋海棠科秋海棠属植物,是球根海棠(B.Tuberons)和野生秋海棠的杂交种。丽格海棠及其亲本球根海棠是目前国际上非常流行的商业化盆花种类。目前,高品质的丽格海棠盆花仍以进口为主,但由于丽格海棠的独特的生理特性造成盆花不耐运输,而且极易造成病原菌从伤口感染,导致品质严重下降。同时丽格海棠抗病性弱,极易感染多种病害,栽培难度大,因此加强丽格海棠品种的遗传改良和抗病生物学研究,进一步深化繁殖和栽培技术研究势在必行。我国植物资源丰富,多种植物富含抗菌有效成分,进行生物农药的开发优势明显。如果能把农业有害杂草开发成生物农药,则是变害为利,将会大大的推动我国植物源农药的发展。
本研究以丽格海棠为材料,通过茎腐病病原菌的分离和鉴定,明确了引起本地丽格海棠茎腐病的病原,在此基础上进行了EMS诱导的抗茎腐病突变体的筛选,壳聚糖生物肥料防治该病的初步研究,筛选了一部分具有高效抑制该种病原的植物种类,并对植物源农药防治丽格海棠茎腐病进行了一些初步研究,为丽格海棠植物的遗传改良和抗病生物学研究奠定了基础。研究结果如下:
1、采用柯赫氏法则和常规组织分离法相结合,分离鉴定出引起丽格海棠茎腐病的病原菌,为丝核菌中的立枯丝核菌(R.solani Kuhn),为后期的抗病突变体的筛选和抗病植物提取液的筛选奠定了基础。
2、对EMS诱导的突变体进行了抗立枯丝核菌引起的茎腐病筛选,经两轮筛选在接种量为3片/株选择压下最终得到3株突变株,发病级数为0级,无明显病害特征出现。并且也产生了一定的形态变异,叶片颜色加深,叶脉变浅、变细,花朵变大,花瓣雄蕊化,花期改变。
3、研究了壳聚糖肥料在提高丽格海棠的园艺性状和抗病性方面的作用,实验结果表明:壳聚糖和NPK的比例为36.3∶13.8∶18.8∶10时,在花期、花朵直径和开花数,POD在40d时酶活,PAL40d时比活力与其他处理有显著差异或与最高值的处理无显著差异,CAT和SOD活性在3个测定时期与其他处理无显著差异,茎腐病和灰霉病的发病率最低,分别为7.14%和0。
4、对15种植物进行了超声波提取,研究了提取液对立枯丝核菌的室内抑制效果,结果筛选到3种中草药具有强烈的抑菌效果,分别是丁香、牡丹皮、花椒。在0.375g干样/100 mL浓度下,三者的抑菌率分别为90.59%、71.76%、55.29%。特别是丁香在0.125 g干样/100 mL浓度下仍然有80.25%的抑菌率,同时其他植物的抑菌率大大下降,并且很多种类表现出了负抗性。
Rieger begonia, whose scientific name is Begonia×hiemalis and synonym rose begonia, belongs to Begoniaceae. It's a hybrid species crossed by B.Tuberons and wild begonia. Rieger begonia and its parent B.Tuberons. as business pot flowers are very popular in the world. High qualitative Rieger begonia is still imported recently, but for its distinct physiological nature, it's difficult to transport in long time. The quality gets worse for germs infecting the wound easily, while the disease resistance is poor. So it's necessary extremely to enhance genetic improvement, study the pathology and deepen the breed and culture technique. Our country is rich in plant resource and many kinds of them have effective component to resist disease, which is the superiority to develop. This development will advance the development of Biotanical biochemical pesticides if we turn agriculture harmful weed to Biological Pesticide.
The study settle Rieger begonia as material, through isolating and identifying stem rotten pathogen, clear-cut the local pathogen of Rieger Begonia from the base of stem. We bolt the the mutants induced by EMS to resist stem rotten, prevent and cure the rotten by chitosan biofertilizer and bolt some plants with effective component to resist this disease, study preliminary biotanical biochemical pesticides. These studies establish the base of genctic improvement and pathology of Rieger begonia. The general results are following:
1. Isolating and identifying stem rotten pathogen of Rieger Begonia, settle the base of later stage to bolt the mutant and plant extract of resisting stem rotten. We integrated the Koch's Postulates and common tissue separation, isolated the pathogen R. solani Kuhn.
2. Screening the mutants induced by EMS to resist stem rotten of Rieger begonia, after two turns selection, three mutants were got which can resist the R. solani Kuhn. They are selected under the inoculum of three pills per pot with disease progression zero, which is no obvious disorder signature. Besides the mutants have some morphosis variation including these leaves turn greener, vein turns lighter and thinner, flower turns larger, petals turn to stamen and florescence changes.
3. Studying the effects of chitosan biofertilizer increasing horticultural characters and the disease resistance, the results demonstrate: florescence, flower diameter and quantity, the activity of POD and PAL in 40d under chitosan and NPK at ratio 36.3:13.8:18.8:10 display significant deviation compared with others or display insignificant deviation compared with the highest treatment while CAT and SOD with indistinctive deviation in the three determination period. The treatment has the lowest incidence of stem rotten and gray mold, which is 7.14% and 0 respectively.
4. Extracting 15 plants with ultrasonic wave, study if the extract inhibit the R. solani Kuhn ex vivo. As the results, we got three Chinese herbal medicine which have the intensive bacteriostasis effect, they are caryophyllus, Japan pepper and tree peony bark. Under the concentration 0.375 g/100mL ,their inhibition ratio is 90.59%、71.76% and 55.29% respectively. Especially caryophyllus still inhibit R. solani Kuhn strong under the concentration 0.125g/100mL with the inhibition ratio 80.25% while many others' ratio drop consumedly and display the negative resistance.
本研究以丽格海棠为材料,通过茎腐病病原菌的分离和鉴定,明确了引起本地丽格海棠茎腐病的病原,在此基础上进行了EMS诱导的抗茎腐病突变体的筛选,壳聚糖生物肥料防治该病的初步研究,筛选了一部分具有高效抑制该种病原的植物种类,并对植物源农药防治丽格海棠茎腐病进行了一些初步研究,为丽格海棠植物的遗传改良和抗病生物学研究奠定了基础。研究结果如下:
1、采用柯赫氏法则和常规组织分离法相结合,分离鉴定出引起丽格海棠茎腐病的病原菌,为丝核菌中的立枯丝核菌(R.solani Kuhn),为后期的抗病突变体的筛选和抗病植物提取液的筛选奠定了基础。
2、对EMS诱导的突变体进行了抗立枯丝核菌引起的茎腐病筛选,经两轮筛选在接种量为3片/株选择压下最终得到3株突变株,发病级数为0级,无明显病害特征出现。并且也产生了一定的形态变异,叶片颜色加深,叶脉变浅、变细,花朵变大,花瓣雄蕊化,花期改变。
3、研究了壳聚糖肥料在提高丽格海棠的园艺性状和抗病性方面的作用,实验结果表明:壳聚糖和NPK的比例为36.3∶13.8∶18.8∶10时,在花期、花朵直径和开花数,POD在40d时酶活,PAL40d时比活力与其他处理有显著差异或与最高值的处理无显著差异,CAT和SOD活性在3个测定时期与其他处理无显著差异,茎腐病和灰霉病的发病率最低,分别为7.14%和0。
4、对15种植物进行了超声波提取,研究了提取液对立枯丝核菌的室内抑制效果,结果筛选到3种中草药具有强烈的抑菌效果,分别是丁香、牡丹皮、花椒。在0.375g干样/100 mL浓度下,三者的抑菌率分别为90.59%、71.76%、55.29%。特别是丁香在0.125 g干样/100 mL浓度下仍然有80.25%的抑菌率,同时其他植物的抑菌率大大下降,并且很多种类表现出了负抗性。
Rieger begonia, whose scientific name is Begonia×hiemalis and synonym rose begonia, belongs to Begoniaceae. It's a hybrid species crossed by B.Tuberons and wild begonia. Rieger begonia and its parent B.Tuberons. as business pot flowers are very popular in the world. High qualitative Rieger begonia is still imported recently, but for its distinct physiological nature, it's difficult to transport in long time. The quality gets worse for germs infecting the wound easily, while the disease resistance is poor. So it's necessary extremely to enhance genetic improvement, study the pathology and deepen the breed and culture technique. Our country is rich in plant resource and many kinds of them have effective component to resist disease, which is the superiority to develop. This development will advance the development of Biotanical biochemical pesticides if we turn agriculture harmful weed to Biological Pesticide.
The study settle Rieger begonia as material, through isolating and identifying stem rotten pathogen, clear-cut the local pathogen of Rieger Begonia from the base of stem. We bolt the the mutants induced by EMS to resist stem rotten, prevent and cure the rotten by chitosan biofertilizer and bolt some plants with effective component to resist this disease, study preliminary biotanical biochemical pesticides. These studies establish the base of genctic improvement and pathology of Rieger begonia. The general results are following:
1. Isolating and identifying stem rotten pathogen of Rieger Begonia, settle the base of later stage to bolt the mutant and plant extract of resisting stem rotten. We integrated the Koch's Postulates and common tissue separation, isolated the pathogen R. solani Kuhn.
2. Screening the mutants induced by EMS to resist stem rotten of Rieger begonia, after two turns selection, three mutants were got which can resist the R. solani Kuhn. They are selected under the inoculum of three pills per pot with disease progression zero, which is no obvious disorder signature. Besides the mutants have some morphosis variation including these leaves turn greener, vein turns lighter and thinner, flower turns larger, petals turn to stamen and florescence changes.
3. Studying the effects of chitosan biofertilizer increasing horticultural characters and the disease resistance, the results demonstrate: florescence, flower diameter and quantity, the activity of POD and PAL in 40d under chitosan and NPK at ratio 36.3:13.8:18.8:10 display significant deviation compared with others or display insignificant deviation compared with the highest treatment while CAT and SOD with indistinctive deviation in the three determination period. The treatment has the lowest incidence of stem rotten and gray mold, which is 7.14% and 0 respectively.
4. Extracting 15 plants with ultrasonic wave, study if the extract inhibit the R. solani Kuhn ex vivo. As the results, we got three Chinese herbal medicine which have the intensive bacteriostasis effect, they are caryophyllus, Japan pepper and tree peony bark. Under the concentration 0.375 g/100mL ,their inhibition ratio is 90.59%、71.76% and 55.29% respectively. Especially caryophyllus still inhibit R. solani Kuhn strong under the concentration 0.125g/100mL with the inhibition ratio 80.25% while many others' ratio drop consumedly and display the negative resistance.
引文
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