超声+热处理对猕猴桃枝条组织中溃疡病菌的杀菌作用及对苗木生长的影响
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摘要
【目的】猕猴桃溃疡病是由Pseudomonas syringae pv.actinidiae(Psa)引起的毁灭性细菌性病害,为杀灭猕猴桃枝条组织中的Psa,研究超声+热处理对猕猴桃枝条组织中Psa的杀菌作用及对苗木生长的影响。【方法】通过单因素试验温度、超声频率及处理时间,以杀菌率为检测指标,使用响应面分析法优化杀菌工艺并进一步验证,在此基础上处理猕猴桃幼苗,观察其成活率、叶片面积、苗高、叶片数及地际直径的生长情况。【结果】研究结果表明,最佳杀菌工艺条件为:温度45℃,超声频率80 KHz,时间30 min,此条件下杀菌率为97.75%。同时,在最佳工艺条件下,处理猕猴桃幼苗,与对照组相比,处理组苗木全部成活,且其叶片面积、苗高、叶片数、地际直径无显著差异。【结论】温度45℃,超声频率80 KHz,时间30 min处理既能杀灭猕猴桃枝条组织、幼苗中的Psa,又不影响其正常生长,为控制猕猴桃苗木、枝条传播溃疡病菌提供了理论基础。
【Objective】Bacterial canker of kiwifruit caused by Pseudomonas seudomonas syringae pv.actinidiae(Psa) is currently the major threat to its commercial production worldwide, which belongs to a typical low-temperature disease and mainly infects the arms, lateral branches and leaves of kiwifruit.The main transmission routes of P. syringae pv. actinidiae(Psa) who always has colonized in the phloem and xylem ducts are boughs and seedlings. As traditional chemical preventions are difficult to kill P.syringae pv. actinidiae(Psa) parasitizing in branches, nursery stock treatment is frequently more effective. Thus, the effects of ultrasonic + heat treatment on killing P. syringae pv. actinidiae(Psa) and seedling growth are evaluated, respectively, which can provide a theoretical basis for bacterial canker controlling by reducing the spread of P. syringae pv. actinidiae(Psa) in kiwifruit branches and seedlings.【Methods】Kiwifruit branches with bacterial canker were selected as raw materials, and the sterilization rates of ultrasonic + heat treatment were determinated through the colony counting method. Single factor experiment was carried out at different treatment temperatures(6 gradients every 5 ℃ from 25 to50 ℃), treatment times(7 gradients every 10 min from 0 to 60 min), and ultrasonic frequencies(6 gradients every 20 kHz from 0 to 100 kHz). Based on the optimum treatment temperature, ultrasonic frequency and treatment time were obtained from the single factor test, and then Box-Benhnken center experiment(Taking the best value point of each single factor as the center and from the upper and lower regions taking one level value as the design level, 17 test points were designed with three factors and three levels, among which the zero point test was repeated 5 times to estimate the test error), response surface and the Design – Expert 8.0.5 b statistical software regression analysis were used to obtain optimum sterilization process after ultrasonic + heat treatment. Finally, kiwifruit seedlings treated by the optimum process were planted in field to observe their survival rate, leaf area, seedling height, leaf number and growth situation of near-ground diameter.【Results】The best single factor condition was treating temperature at 45 ℃, ultrasonic frequency at 80 kHz and treating time for 30 min, respectively. Further response surface analysis showed that the sterilization rate was up to 97.75% after the optimal sterilization process was applied(treating temperature at 45 ℃, ultrasonic frequency at 80 kHz and treatment time for 30 min). The results of regression analysis briefly showed that the sterilization rate under optimal conditions was 97.88%, which showed no significant difference from the results of response surface optimization(α = 0.01). The results of model variance analysis showed that the regression model was very significant(p = 0.000 1), and the lack of fit(p = 0.083 3 > 0.01) showed no significant difference, which meant that the fitting degree of the model had the greatest influence. And following model variance analysis showed that both the primary terms( A(p < 0.000 1)and C(p = 0.000 3) and secondary terms(A2(p < 0.000 1)and C2(p = 0.000 7)) had a significant difference in the model. And the A and A2 were very significant, which indicated that temperature was the most important factor in the sterilization process. Besides, the interaction showed that the order of interaction strength was AC >BC > AB, which meant the interaction between treatment temperature and time had the greatest influence on the sterilization rate. Furthermore, the effect of each single factor on sterilization rate hardly showed a simple linear relationship. The order of the efficacy is A(treatment temperature) > B(treatment time) > C(ultrasonic frequency). In the end, the results of transplanting experiment of the treated kiwifruit plants showed that the survival rate of the treated group was 100%, and the leaf area, seedling height, number of leaves and near-ground diameter had no significant difference between the control and treatment(α = 0.05).【Conclusion】The results remarkably showed that the optimal sterilization process of ultrasonic and heat treatment(treating temperature at 45 ℃, ultrasonic frequency at 80 kHz and treating time for 30 min)can not only kill P. syringae pv. actinidiae(Psa) living in kiwifruit shoots and seedlings effectively, but also have no negative effect on its survival rate and growth. And this method was clean, efficient, convenient and quick, comparing with the current chemical and biological prevention. What's more, this treatment provided a new direction in the prevention and treatment of P. syringae pv. actinidiae(Psa). Therefore, this study has a potential application for the P. syringae pv. actinidiae(Psa) controlling in the infected branches of kiwifruit plantings in the field.
【Objective】Bacterial canker of kiwifruit caused by Pseudomonas seudomonas syringae pv.actinidiae(Psa) is currently the major threat to its commercial production worldwide, which belongs to a typical low-temperature disease and mainly infects the arms, lateral branches and leaves of kiwifruit.The main transmission routes of P. syringae pv. actinidiae(Psa) who always has colonized in the phloem and xylem ducts are boughs and seedlings. As traditional chemical preventions are difficult to kill P.syringae pv. actinidiae(Psa) parasitizing in branches, nursery stock treatment is frequently more effective. Thus, the effects of ultrasonic + heat treatment on killing P. syringae pv. actinidiae(Psa) and seedling growth are evaluated, respectively, which can provide a theoretical basis for bacterial canker controlling by reducing the spread of P. syringae pv. actinidiae(Psa) in kiwifruit branches and seedlings.【Methods】Kiwifruit branches with bacterial canker were selected as raw materials, and the sterilization rates of ultrasonic + heat treatment were determinated through the colony counting method. Single factor experiment was carried out at different treatment temperatures(6 gradients every 5 ℃ from 25 to50 ℃), treatment times(7 gradients every 10 min from 0 to 60 min), and ultrasonic frequencies(6 gradients every 20 kHz from 0 to 100 kHz). Based on the optimum treatment temperature, ultrasonic frequency and treatment time were obtained from the single factor test, and then Box-Benhnken center experiment(Taking the best value point of each single factor as the center and from the upper and lower regions taking one level value as the design level, 17 test points were designed with three factors and three levels, among which the zero point test was repeated 5 times to estimate the test error), response surface and the Design – Expert 8.0.5 b statistical software regression analysis were used to obtain optimum sterilization process after ultrasonic + heat treatment. Finally, kiwifruit seedlings treated by the optimum process were planted in field to observe their survival rate, leaf area, seedling height, leaf number and growth situation of near-ground diameter.【Results】The best single factor condition was treating temperature at 45 ℃, ultrasonic frequency at 80 kHz and treating time for 30 min, respectively. Further response surface analysis showed that the sterilization rate was up to 97.75% after the optimal sterilization process was applied(treating temperature at 45 ℃, ultrasonic frequency at 80 kHz and treatment time for 30 min). The results of regression analysis briefly showed that the sterilization rate under optimal conditions was 97.88%, which showed no significant difference from the results of response surface optimization(α = 0.01). The results of model variance analysis showed that the regression model was very significant(p = 0.000 1), and the lack of fit(p = 0.083 3 > 0.01) showed no significant difference, which meant that the fitting degree of the model had the greatest influence. And following model variance analysis showed that both the primary terms( A(p < 0.000 1)and C(p = 0.000 3) and secondary terms(A2(p < 0.000 1)and C2(p = 0.000 7)) had a significant difference in the model. And the A and A2 were very significant, which indicated that temperature was the most important factor in the sterilization process. Besides, the interaction showed that the order of interaction strength was AC >BC > AB, which meant the interaction between treatment temperature and time had the greatest influence on the sterilization rate. Furthermore, the effect of each single factor on sterilization rate hardly showed a simple linear relationship. The order of the efficacy is A(treatment temperature) > B(treatment time) > C(ultrasonic frequency). In the end, the results of transplanting experiment of the treated kiwifruit plants showed that the survival rate of the treated group was 100%, and the leaf area, seedling height, number of leaves and near-ground diameter had no significant difference between the control and treatment(α = 0.05).【Conclusion】The results remarkably showed that the optimal sterilization process of ultrasonic and heat treatment(treating temperature at 45 ℃, ultrasonic frequency at 80 kHz and treating time for 30 min)can not only kill P. syringae pv. actinidiae(Psa) living in kiwifruit shoots and seedlings effectively, but also have no negative effect on its survival rate and growth. And this method was clean, efficient, convenient and quick, comparing with the current chemical and biological prevention. What's more, this treatment provided a new direction in the prevention and treatment of P. syringae pv. actinidiae(Psa). Therefore, this study has a potential application for the P. syringae pv. actinidiae(Psa) controlling in the infected branches of kiwifruit plantings in the field.
引文
[1]EVERETT K R,TAYLOR R K,ROMBERG M K,REES-GEORGE J,FULLERTON R A,VANNESTE J L.First report of Pseudomonas syringae pv.actinidiae causing kiwifruit bacterial canker in New Zealand[J].Australisian Plant Disease Notes,2011,6(1):67-71.
[2]ABELLEIRA A,LóPEZ M M,PE?ALVER J,AGUíN O,MANSILLA J P,PICOAGA A.First report of bacterial canker of kiwifruit can used by Pseudomonas syringae pv.actinidiae in Spain[J].Plant Disease,2011,95(12):1583-1583.
[3]SCORTICHINI M.Occurrence of Pseudomonas syringae pv.actinidiae on kiwifruit in Italy[J].Plant Pathology,1994,43(6):1035-1038.
[4]BASTAS K K,KARAKAYA A.First report of bacterical canker of kiwifruit caused by Pseudomonas syringae pv.actinidiae in Turkey[J].Plant Disease,2012,96(3):452-452.
[5]胡家勇.猕猴桃溃疡病菌株收集及其寄主和传播媒介的鉴定[D].合肥:安徽农业大学,2014.HU jiayong.Collection of kiwi canker strains and identification of their host and vectors identification[D].Hefei:Anhui Agricultural University,2014.
[6]李亚巍,王小洁,吴群,李士谣,李双荣,朱立武,刘普.猕猴桃溃疡病菌的田间分布及其传播规律[J/OL].生物学杂志[2018-07-26].http://kns.cnki.net/kcms/detail/34.1081.Q.20180726.15-46.012.html.LI Yawei,WANG Xiaojie,WU Qun,LI Shiyao,LI Shuangrong,ZHU Liwu,LIU Pu.The field distribution and prevalence rules of Pseudomonas syringae pv.actinidiae in kiwifruit[J/OL].Journal of Biology[2018-07-26].http://kns.cnki.net/kcms/detail/34.-1081.Q.20180726.1546.012.html.
[7]黄其玲,高小宁,赵志博,秦虎强,黄丽丽.GFPuv标记猕猴桃溃疡病菌的生物学特性及其在土壤、根系中的定殖[J].中国农业科学,2013,46(2):282-291.HUANG Qiling,GAO Xiaoning,ZHAO Zhibo,QIN Huqiang,HUANG Lili.Biological characteristics of GFPuv-labeled kiwi canker and its colonization in soil and roots of kiwifruit[J].Scientia Agricultura Sinica,2013,46(2):282-291.
[8]MAZZAGLIA A,STUDHOLME D J,TARATUFOLO M C,CAIL R,ALMEDLA N F.Pseudomonas syringae pv.actinidiae(PSA)isolates from recent bacterial canker of kiwifruit outbreaks belong to the same genetic lineage[J].PLoS One,2012,7(5):e36518.
[9]TAKIKAWA Y,SERIZAWAE S,ICHIKAWA T,TSUYUMU S,GOTO M.Pseudomonas syringae pv.nov.the causal bacterium of canker of kiwifruit in Japan[J].Annals of the Phytopathologicial Society of Japan,1989,55(4):437-444.
[10]黄其玲.应用GFPuv标记技术研究猕猴桃溃疡病菌在组织中的侵染扩展动态[D].杨凌:西北农林科技大学,2013.HUANG Qiling.Use of the green fluorescent protein variant GF-Puv for analysis of Pseudomonas syringae pv.actinidiae infection,movement and colonization in kiwifruit[D].Yangling:Northwest A&F University,2013.
[11]陈虹君,杨国安,迟旭春.猕猴桃溃疡病致病因素与防治经验探讨[J].南方农业,2018,12(17):1-2.CHEN Hongjun,YANG Guo’an,CHI Xuchun.Pathogenic factors and prevention experience of kiwifruit ulcer disease[J].South China Agriculture,2018,12(17):1-2.
[12]朱海云,马瑜,柯杨,李勃,孙超.猕猴桃细菌性溃疡病生防菌的筛选、鉴定及其防效初探[J].微生物学杂志,2016,36(5):90-96.ZHU Haiyun,MA Yu,KE Yang,LI Bo,SUN Chao.Screening,identification and controlling effects of biocontrol strain against chinese gooseberry or kiwifruit bacterial canker[J].Journal of Microbiology,2016,36(5):90-96.
[13]秦虎强,赵志博,高小宁,杨彪,徐国信,黄丽丽.猕猴桃细菌性溃疡病菌对17种杀菌剂的敏感性及不同药剂田间防效[J].西北农业学报,2015,24(9):145-151.QIN Huqiang,ZHAO Zhibo,GAO Xiaoning,YANG Biao,XUGuoxin,HUANG Lili.Bacteridal activities of sevevteen bactericides and their field applications against Pseudomonas syringae pv.actinidiae[J].Acta Agriculturae Boreali-occidentalis Sinica,2015,24(9):145-151.
[14]HAN H S,NAM H Y,KOH Y J,HUR J S,JUNG J S.Molecularbases of high-level streptomycin resistance in Pseudomonas marginlis and Pseudomonas syringae pv.actinidiae[J].The Journal of Microbiolgy,2003,41(1):16-21.
[15]NAJIMA M,GOTO M,HIBI T.Similarity between copper resistance genes from Pseudomonas syringae pv.actinidiae and P.syringae pv.tomato[J].Journal of General Plant Pathology,2002,68(1):68-74.
[16]余桂容,张敏,叶华智.小麦赤霉病的生物防治研究Ⅰ.拮抗芽孢杆菌的分离、筛选、鉴定和防病效果[J].四川农业大学学报,1998,16(3):28-32.YU Guirong,ZHANG Min,YE Huazhi.Studies on biological control of wheat head blight with BACLLUS SPP.Ⅰ.Isolation,Identification and disease suppressing efficasy of antagonists[J].Journal of Sichuan Agricultural University,1998,16(3):28-32.
[17]林孔湘,骆学海.柑桔黄梢(黄龙)病热治疗的初步研究[J].植物保护学报,1965,3(2):169-175.LIN Kongxiang,LUO Xuehai.Preliminary study on the treatment of citrus yellow shoot(Huanglong)disease[J]Journal of Plant Protection,1965,3(2):169-175.
[18]骆学海,罗志达,唐伟文.柑桔黄龙病热治疗的研究[J].植物保护学报,1981,8(1):47-52.LUO Xuehai,LUO Zhida,TANG Weiwen.Studies on the thermotherapy of citrus yellow shoot disease[J].Journal of Plant Protection,1981,8(1):47-52.
[19]高日霞,郑和睦.柑桔黄龙病治疗试验研究[J].植物保护学报,1981,8(1):53-58.GAO Rixia,ZHENG Hemu.On control of yellow shoot disease of citrus[J].Journal of Plant Protection,1981,8(1):53-58.
[20]赵军嫄,李子富,周瑱,周晓琴,杨鑫,王冬泠.超声波/紫外线一体化反应器用于污水消毒的中试研究[J].环境工程学报,2016,10(11):6185-6189.ZHAO Junyuan,LI Zifu,ZHOU Zhen,ZHOU Xiaoqin,YANGXin,WANG Dongling.Pilot study on ultrasonic/ultraviolet integrated reactor for sewage disinfection[J].Journal of Environmental Sciences,2016,10(11):6185-6189.
[21]马海燕,张慜,孙金才,楼芳琼.生冻大根泥(萝卜泥)中微生物控制的物理方法[J].食品与生物技术学报,2011,30(1):78-83.MA Haiyan,ZHANG Min,SUN Jincai,LOU Fangqiong.Physical method for microbial control in raw frozen root mud(radish puree)[J].Journal of Food Science and Biotechnology,2011,30(1):78-83.
[22]MASON T J,PANIWNYK L,LORIMER J P.The uses of ultrasound in food technology[J].Ultrasonics Sonochemistry,1996,3(3):253-260.
[23]中华人民共和国国家卫生和计划生育委员会.食品安全国家标准食品微生物学检验菌落总数测定:GB4789.2-2016[S].北京:中国标准出版社,2016.National Health and Family Planning Commission.National food safety standard-Food microbiology test Total number of colonies:GB4789.2-2016[S].Beijing:China Standard Press,2016.
[24]龙秀琴,许杰.不同育苗方式对光皮树出苗率及苗木生长情况和质量的影响[J].安徽农业科学,2009,37(5):1986-1987.LONG Xiuqin,XU Jie.Effects of different raising breeding mode on germination rate of cornus wilsoniana and its growth status and quality[J].Journal of Anhui Agricultural Sciences,2009,37(5):1986-1987.
[25]穆立蔷,王振宇,李长斌,丰绪霞.活性保水剂对苗木生长及成活率的影响[J].东北林业大学学报,2004(02):19-21.MU Liqiang,WANG Zhengyu,LI Changbin,FENG Xuxia.The effect of bioactive water retaining agent on seeding growth and survival rate[J].Journal of Northeast Forestry University,2004,32(2):19-21.
[26]李泉厂,陈金焕,王西红.不同药剂防治猕猴桃溃疡病效果研究[J].现代农业科技,2013(11):130-131.LI Quanchang,CHEN Jinhuan,WANG Xihong.Study on the effect of different chemicals on prevention and treatment of kiwifruit ulcer[J].Modern Agricultural Science and Technology,2013(11):130-131.
[27]秦虎强,高小宁,赵志博,朱穗层,李建民,黄丽丽.陕西猕猴桃细菌性溃疡病田间发生动态和规律[J].植物保护学报,2013,40(3):225-230.QIN Huqiang,GAO Xiaoning,ZHAO Zhibo,ZHU Suiceng,LIJianming,HUANG Lili.The prevalence dynamics and rules of bacterial canker of kiwifruit in shaanxi[J].Journal of Plant Protection,2013,40(3):225-230.
[28]林孔湘.柑桔黄梢(黄龙)病的进一步研究[J].植物保护学报,1963,1(3):243-251.LIN Kongxiang.Further study on citrus yellow shoot(Huanglong)disease[J].Journal of Plant Protection,1963,1(3):243-251.
[29]贾志成,郑加强,黄雅杰,周宏平,Ehsani Reza.柑橘黄龙病热处理防治技术研究进展[J].农业工程学报,2015,31(23):1-9.JIA Zhicheng,ZHENG Jiaqiang,HUANG Yajie,ZHOU Hongping,EHSANI R.Research progress on heat treatment technology of citrus Huanglongbing[J].Transactions of the Chinese Society of Agricultural Engineering,2015,31(23):1-9.
[30]陈荣风,薛广波,顾春英.超声波消毒研究进展[J].上海预防医学,1999(11):491-495.CHEN Rongfeng,XUE Guangbo,GU Chunying.Progress in ultrasonic disinfection research[J].Shanghai Journal of Preventive Medicine,1999(11):491-495.
[2]ABELLEIRA A,LóPEZ M M,PE?ALVER J,AGUíN O,MANSILLA J P,PICOAGA A.First report of bacterial canker of kiwifruit can used by Pseudomonas syringae pv.actinidiae in Spain[J].Plant Disease,2011,95(12):1583-1583.
[3]SCORTICHINI M.Occurrence of Pseudomonas syringae pv.actinidiae on kiwifruit in Italy[J].Plant Pathology,1994,43(6):1035-1038.
[4]BASTAS K K,KARAKAYA A.First report of bacterical canker of kiwifruit caused by Pseudomonas syringae pv.actinidiae in Turkey[J].Plant Disease,2012,96(3):452-452.
[5]胡家勇.猕猴桃溃疡病菌株收集及其寄主和传播媒介的鉴定[D].合肥:安徽农业大学,2014.HU jiayong.Collection of kiwi canker strains and identification of their host and vectors identification[D].Hefei:Anhui Agricultural University,2014.
[6]李亚巍,王小洁,吴群,李士谣,李双荣,朱立武,刘普.猕猴桃溃疡病菌的田间分布及其传播规律[J/OL].生物学杂志[2018-07-26].http://kns.cnki.net/kcms/detail/34.1081.Q.20180726.15-46.012.html.LI Yawei,WANG Xiaojie,WU Qun,LI Shiyao,LI Shuangrong,ZHU Liwu,LIU Pu.The field distribution and prevalence rules of Pseudomonas syringae pv.actinidiae in kiwifruit[J/OL].Journal of Biology[2018-07-26].http://kns.cnki.net/kcms/detail/34.-1081.Q.20180726.1546.012.html.
[7]黄其玲,高小宁,赵志博,秦虎强,黄丽丽.GFPuv标记猕猴桃溃疡病菌的生物学特性及其在土壤、根系中的定殖[J].中国农业科学,2013,46(2):282-291.HUANG Qiling,GAO Xiaoning,ZHAO Zhibo,QIN Huqiang,HUANG Lili.Biological characteristics of GFPuv-labeled kiwi canker and its colonization in soil and roots of kiwifruit[J].Scientia Agricultura Sinica,2013,46(2):282-291.
[8]MAZZAGLIA A,STUDHOLME D J,TARATUFOLO M C,CAIL R,ALMEDLA N F.Pseudomonas syringae pv.actinidiae(PSA)isolates from recent bacterial canker of kiwifruit outbreaks belong to the same genetic lineage[J].PLoS One,2012,7(5):e36518.
[9]TAKIKAWA Y,SERIZAWAE S,ICHIKAWA T,TSUYUMU S,GOTO M.Pseudomonas syringae pv.nov.the causal bacterium of canker of kiwifruit in Japan[J].Annals of the Phytopathologicial Society of Japan,1989,55(4):437-444.
[10]黄其玲.应用GFPuv标记技术研究猕猴桃溃疡病菌在组织中的侵染扩展动态[D].杨凌:西北农林科技大学,2013.HUANG Qiling.Use of the green fluorescent protein variant GF-Puv for analysis of Pseudomonas syringae pv.actinidiae infection,movement and colonization in kiwifruit[D].Yangling:Northwest A&F University,2013.
[11]陈虹君,杨国安,迟旭春.猕猴桃溃疡病致病因素与防治经验探讨[J].南方农业,2018,12(17):1-2.CHEN Hongjun,YANG Guo’an,CHI Xuchun.Pathogenic factors and prevention experience of kiwifruit ulcer disease[J].South China Agriculture,2018,12(17):1-2.
[12]朱海云,马瑜,柯杨,李勃,孙超.猕猴桃细菌性溃疡病生防菌的筛选、鉴定及其防效初探[J].微生物学杂志,2016,36(5):90-96.ZHU Haiyun,MA Yu,KE Yang,LI Bo,SUN Chao.Screening,identification and controlling effects of biocontrol strain against chinese gooseberry or kiwifruit bacterial canker[J].Journal of Microbiology,2016,36(5):90-96.
[13]秦虎强,赵志博,高小宁,杨彪,徐国信,黄丽丽.猕猴桃细菌性溃疡病菌对17种杀菌剂的敏感性及不同药剂田间防效[J].西北农业学报,2015,24(9):145-151.QIN Huqiang,ZHAO Zhibo,GAO Xiaoning,YANG Biao,XUGuoxin,HUANG Lili.Bacteridal activities of sevevteen bactericides and their field applications against Pseudomonas syringae pv.actinidiae[J].Acta Agriculturae Boreali-occidentalis Sinica,2015,24(9):145-151.
[14]HAN H S,NAM H Y,KOH Y J,HUR J S,JUNG J S.Molecularbases of high-level streptomycin resistance in Pseudomonas marginlis and Pseudomonas syringae pv.actinidiae[J].The Journal of Microbiolgy,2003,41(1):16-21.
[15]NAJIMA M,GOTO M,HIBI T.Similarity between copper resistance genes from Pseudomonas syringae pv.actinidiae and P.syringae pv.tomato[J].Journal of General Plant Pathology,2002,68(1):68-74.
[16]余桂容,张敏,叶华智.小麦赤霉病的生物防治研究Ⅰ.拮抗芽孢杆菌的分离、筛选、鉴定和防病效果[J].四川农业大学学报,1998,16(3):28-32.YU Guirong,ZHANG Min,YE Huazhi.Studies on biological control of wheat head blight with BACLLUS SPP.Ⅰ.Isolation,Identification and disease suppressing efficasy of antagonists[J].Journal of Sichuan Agricultural University,1998,16(3):28-32.
[17]林孔湘,骆学海.柑桔黄梢(黄龙)病热治疗的初步研究[J].植物保护学报,1965,3(2):169-175.LIN Kongxiang,LUO Xuehai.Preliminary study on the treatment of citrus yellow shoot(Huanglong)disease[J]Journal of Plant Protection,1965,3(2):169-175.
[18]骆学海,罗志达,唐伟文.柑桔黄龙病热治疗的研究[J].植物保护学报,1981,8(1):47-52.LUO Xuehai,LUO Zhida,TANG Weiwen.Studies on the thermotherapy of citrus yellow shoot disease[J].Journal of Plant Protection,1981,8(1):47-52.
[19]高日霞,郑和睦.柑桔黄龙病治疗试验研究[J].植物保护学报,1981,8(1):53-58.GAO Rixia,ZHENG Hemu.On control of yellow shoot disease of citrus[J].Journal of Plant Protection,1981,8(1):53-58.
[20]赵军嫄,李子富,周瑱,周晓琴,杨鑫,王冬泠.超声波/紫外线一体化反应器用于污水消毒的中试研究[J].环境工程学报,2016,10(11):6185-6189.ZHAO Junyuan,LI Zifu,ZHOU Zhen,ZHOU Xiaoqin,YANGXin,WANG Dongling.Pilot study on ultrasonic/ultraviolet integrated reactor for sewage disinfection[J].Journal of Environmental Sciences,2016,10(11):6185-6189.
[21]马海燕,张慜,孙金才,楼芳琼.生冻大根泥(萝卜泥)中微生物控制的物理方法[J].食品与生物技术学报,2011,30(1):78-83.MA Haiyan,ZHANG Min,SUN Jincai,LOU Fangqiong.Physical method for microbial control in raw frozen root mud(radish puree)[J].Journal of Food Science and Biotechnology,2011,30(1):78-83.
[22]MASON T J,PANIWNYK L,LORIMER J P.The uses of ultrasound in food technology[J].Ultrasonics Sonochemistry,1996,3(3):253-260.
[23]中华人民共和国国家卫生和计划生育委员会.食品安全国家标准食品微生物学检验菌落总数测定:GB4789.2-2016[S].北京:中国标准出版社,2016.National Health and Family Planning Commission.National food safety standard-Food microbiology test Total number of colonies:GB4789.2-2016[S].Beijing:China Standard Press,2016.
[24]龙秀琴,许杰.不同育苗方式对光皮树出苗率及苗木生长情况和质量的影响[J].安徽农业科学,2009,37(5):1986-1987.LONG Xiuqin,XU Jie.Effects of different raising breeding mode on germination rate of cornus wilsoniana and its growth status and quality[J].Journal of Anhui Agricultural Sciences,2009,37(5):1986-1987.
[25]穆立蔷,王振宇,李长斌,丰绪霞.活性保水剂对苗木生长及成活率的影响[J].东北林业大学学报,2004(02):19-21.MU Liqiang,WANG Zhengyu,LI Changbin,FENG Xuxia.The effect of bioactive water retaining agent on seeding growth and survival rate[J].Journal of Northeast Forestry University,2004,32(2):19-21.
[26]李泉厂,陈金焕,王西红.不同药剂防治猕猴桃溃疡病效果研究[J].现代农业科技,2013(11):130-131.LI Quanchang,CHEN Jinhuan,WANG Xihong.Study on the effect of different chemicals on prevention and treatment of kiwifruit ulcer[J].Modern Agricultural Science and Technology,2013(11):130-131.
[27]秦虎强,高小宁,赵志博,朱穗层,李建民,黄丽丽.陕西猕猴桃细菌性溃疡病田间发生动态和规律[J].植物保护学报,2013,40(3):225-230.QIN Huqiang,GAO Xiaoning,ZHAO Zhibo,ZHU Suiceng,LIJianming,HUANG Lili.The prevalence dynamics and rules of bacterial canker of kiwifruit in shaanxi[J].Journal of Plant Protection,2013,40(3):225-230.
[28]林孔湘.柑桔黄梢(黄龙)病的进一步研究[J].植物保护学报,1963,1(3):243-251.LIN Kongxiang.Further study on citrus yellow shoot(Huanglong)disease[J].Journal of Plant Protection,1963,1(3):243-251.
[29]贾志成,郑加强,黄雅杰,周宏平,Ehsani Reza.柑橘黄龙病热处理防治技术研究进展[J].农业工程学报,2015,31(23):1-9.JIA Zhicheng,ZHENG Jiaqiang,HUANG Yajie,ZHOU Hongping,EHSANI R.Research progress on heat treatment technology of citrus Huanglongbing[J].Transactions of the Chinese Society of Agricultural Engineering,2015,31(23):1-9.
[30]陈荣风,薛广波,顾春英.超声波消毒研究进展[J].上海预防医学,1999(11):491-495.CHEN Rongfeng,XUE Guangbo,GU Chunying.Progress in ultrasonic disinfection research[J].Shanghai Journal of Preventive Medicine,1999(11):491-495.