芒果果实潜伏侵染、Botryodiplodia theobromae致腐机理及蒂腐病防治技术基础研究
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摘要
芒果(Mangifera indica L.)是一种重要的热带亚热带水果,芒果产业已构成我国热区农业和农村经济的重要组成部分,在热区农业产业结构调整中发挥着积极的作用。然而,芒果采后病害严重制约着芒果产业的发展。由于引起芒果采后病害的病原菌大多具有潜伏侵染的特性,导致果实在贮运及销售过程中快速、严重腐烂,其中,由可可球二孢菌(Botryodiplodia theobromae Pat.)等引起的蒂腐病在果实尚未成熟即开始造成腐烂,是引起芒果采后腐烂的重要病原菌。
本文以可可球二孢菌(B. theobromae)等蒂腐病病菌和芒果互作为研究对象,开展芒果果实真菌潜伏侵染、B. theobromae在果实致病过程中的作用机理、B. theobromae对常用杀菌剂的抗性及其引起的采后病害防治技术等研究,旨在摸清我国芒果果实中的潜伏侵染真菌种类及严重程度,初步阐明B. theobromae致芒果果实快速腐烂的机理,探讨B. theobromae的抗药性水平及蒂腐病的安全、有效的采后处理方法,为研发芒果蒂腐病的防治措施提供依据。
本研究采用组织分离法对采自我国6省区的芒果果实潜伏侵染真菌进行检测与种类鉴定,研究结果表明：在芒果果实中普遍存在真菌的潜伏侵染现象。不同省区的带菌率存在显著差异(F=3.59,P=0.0246)。不同年份的带菌率略有差异,但未达到显著差异水平(F=1.57,P=0.2372)。果实不同部位的带菌率间存在极显著差异(F=46.44,P<0.0001),果柄和果实蒂部的带菌率极显著高于果实中部和端部的带菌率。鉴定的潜伏侵染真菌有11种,分别为Colletotrichum gloeosporioides (Penz.) Penz.&Sacc.、 Botryodiplodia theobromae Pat.、Colletotrichum acutatum Simmonds、Phomopsis mangiferae Ahmad.、Pestalotiopsis mangiferae (P. Henn.) Sutton.、Dothiorella dominicana Pet. et Cif.、 Macrophoma mangiferae Hing.、O. P. Sharma、Alternaria alternata (Fr.) Keissl.、 Aspergillus sp.、Curvularia sp.和Fusarrium spp.,其中,C. gloeosporioides和B. theobromae为芒果果实中的主要潜伏侵染真菌,M. mangiferae在芒果果实上作为潜伏侵染真菌为首次报道,B. theobromae为致病力最强的潜伏侵染真菌。
B. theobromae、C. gloeosporioides、D. dominicana和P. mangiferae4种芒果主要蒂腐病菌的致病力存在极显著差异,其中B. theobromae的致病力最强,B. theobromae在含果胶和纤维素的培养基上的生长速度均极显著高于其他3种蒂腐病菌的生长速度,采用刚果红平板水解圈法测定的结果表明,该菌产生的果胶酶活性极显著高于其他3种蒂腐病菌的果胶酶活性；纤维素酶活性虽与C. gloeosporioides无显著差异,但极显著高于D. dominicana和P. mangiferae的纤维素酶活性。采用3,5-二硝基水杨酸(DNS)比色法测定的结果表明,B. theobromae在离体培养条件下或接种与芒果果实上均可产生多聚半乳糖醛酸酶(PG)、多聚半乳糖醛酸反式消除酶(PGTE)、多聚甲基半乳糖醛酸酶(PMG)、果胶甲基反式消除酶(PMTE),以及纤维素酶(Cx)。其中PG、PMG和Cx活性较高,PGTE和PMTE活性很低。在大量产生的3种细胞壁降解酶中,其中PG活性最大,且峰值在早期出现。
B. theobromae在液体培养基中可产生导致芒果嫩叶褐变、嫩枝萎蔫和果实腐烂的致病毒素。该毒素在酸性至中性条件稳定,热稳定性强,对光照不敏感,低温条件更有利于毒素的保存。
B. theobromae在离体条件下可产生乙烯；接种到芒果果实上,在芒果果实内可诱导芒果果肉产生大量的1-氨基环丙烷-1-羧酸(ACC),从而诱导芒果果实快速释放乙烯。自然条件下发生蒂腐病的果实,在症状出现前1-2d,其乙烯释放速率即开始升高,并随着病程的发展,乙烯释放速率显著增强,且3-5d内,发病果实软化、腐烂。
B. theobromae群体中存在着对多菌灵抗性的群体。采用生长速率法进行了23种杀菌剂对4株对多菌灵抗性差异的B. theobromae的毒力测定,通过EC50值、EC90值及与多菌灵的交互抗性等的综合分析,认为咪鲜胺锰盐、咪鲜胺、丙环唑、苯醚甲环唑、氟硅唑、异菌脲、戊唑醇、腈菌唑、吡唑醚菌酯、井冈霉素,以及百菌清和代森锰锌可用作芒果蒂腐病防治的杀菌剂。B. theobromae除对多菌灵产生抗性外,还对甲基硫菌灵、噻菌灵、醚菌酯和烯唑醇4种杀菌剂产生了抗药性。通过交互抗性分析,多菌灵、噻菌灵、甲基硫菌灵和烯唑醇4种杀菌剂之间存在交互抗性,而醚菌酯与井冈霉素之间存在负交互抗性。
异菌脲、咪鲜胺和咪鲜胺·异菌脲3种杀菌剂对芒果蒂腐病具有较好的防效,而咪鲜胺·异菌脲、咪鲜胺对芒果炭疽病的防效达93%以上。根据对芒果蒂腐病和炭疽病防治效果的综合评价,咪鲜胺·异菌脲对控制芒果采后病害的效果最好,其次是咪鲜胺。
供试的18种食品添加剂对C. gloeosporioidesS、C. gloeosporioidesR、B.theobromaesS、 B. theobromaeR、P. mangiferae和D. dominicana6种芒果采后病害病原菌的毒力存在极显著差异,邻苯基苯酚钠对6种病原菌的平均ECso值和平均EC90值均最小,分别为7.21μg/mL和39.53μg/mL,其次是脱氢乙酸钠和阿魏酸。脱氢乙酸钠对炭疽病和蒂腐病的防效最好,防效分别达到83.52%和59.32%,其次阿魏酸和肉桂醛。阿魏酸处理的果实贮藏时间最长,平均贮藏时间达15.51d,其次是脱氢乙酸钠和甘草抗氧化剂,而毒力最大的邻苯基苯酚钠对芒果果实造成伤害。
使用食品添加剂结合壳聚糖的复合涂膜技术,可显著提高对蒂腐病的防治效果,1000μg/mL脱氢乙酸钠+1.5%壳聚糖涂膜与1000μg/mL阿魏酸+1.5%壳聚糖涂膜,对蒂腐病的防效分别达到99.34%和96.97%,500μg/mL脱氢乙酸钠+1.5%壳聚糖的防效达到90.28%。食品添加剂的复合涂膜技术不仅提高了对芒果蒂腐病的防治效果,还可显著降低杀菌剂的使用,且不存在农药残留问题,是一种值得推广应用的采后处理方法。
Mango (Mangifera indica Linn.), a valuable fruit crop growing throughout the tropics. However, mango is highly susceptible to postharvest decay during storage and transportation, which leads to huge economic losses. The pathogens can infect the mango flowers and immature fruit and remain quiescent until storage, during which the lesions progressively appear and symptoms become more severe along with the extension of time. It has been well known that the stem-end rot caused by Botryodiplodia theobromae is the one of most destructive postharvest disease in mango fruit.
Using B. theobromae and mango fruit as interaction system in this study, we investigated the occurrence in latent infection of pathogenic fungi in six provinces of China according to the tissue isolation method, explored the pathogenic mechanism of B. theobromae, studied the tolerance of B. theobromae to several common fungicides as well as tested some technologies to control postharvest disease in mango. The results were presented as follows:
The results showed that the latent infection of fungi comprehensively existed in the healthy mango fruits. There were significant differences in latent infection rate among the mango fruits in different provinces (F=3.59, P=0.0246), but not significant differences in latent infection rate in different years(F=1.57, P=0.2372). The rates of latent infection in the shoulder and the stem end were significantly higher than those in the other parts (F=46.44, P <0.0001). There were11pathogenic fungi to be identified, including Colletotrichum gloeosporioides (Penz.) Penz.&Sacc、Botryodiplodia theobromae Pat.、Colletotrichum acutatum Simmonds、Phomopsis mangiferae Ahmad.、Pestalotiopsis mangiferae (P. Henn.) Sutton.、Dothiorella dominicana Pet. et Cif、Macrophoma mangiferae Hing.&O. P. Sharma、 Alternaria alternata(Fr.)Keissl.、Aspergillus sp.、Curvularia sp. and Fusarrium spp.. In these isolated fungi, both C. gloeosporioides and B. theobromae were the main fungi leading to latent infection in mango. Moreover, M. mangiferae was first reported as one of latent infection fungi in mango fruit.
Four tested pathogens including B. theobromae, C. gloeosporioides, D. dominicana and P. mangiferae had different pathogenicity that cause stem end rot in mango fruit, among which the B. theobromae showed the strongest pathogenicity. The growth rate of B. theobromae in culture medium containing pectin or cellulose was extremely higher than that in other three pathogens. Furthermore, the tested results using congo red staining method showed that the activity of pectinase generated by B. theobromae was higher than that in other three pathogens, and cellulase activity produced by B. theobromae was higher than that in D. dominicana and P. mangiferae, but was similar as gloeosporioides. The results by colorimetric method of DNS demonstrated that B. theobromae may release polygalacturonase (PG), pectinmethylgalacturonase (PMG), polygalacturonic acid trans-eliminase (PGTE), pectin methyltrans-eliminase (PMTE) and cellulase (Cx.) when cultured in vitro or in vivo. Among three primary CWDEs, PG and Cx appeared activity peak earlier and activity peak of PMG was lagged.
B. theobromae in PA liquid culture produced a specific toxin that can result in browning of mango young leaves, wilting in branches and decay in fruits. The toxin had a strong thermal and acid-tolerance stability but not sensitive to light. The appropriate low temperature was favorable to toxin preservation.
B. theobromae produced ethylene in vitro, and it induced the mango fruit to produce large quantities of1-aminocyclopropane-1-carboxylic acid (ACC), which might be responsible for ethylene evolution. The ethylene began to generate increased prior to appearance in symptom of stem-end rot, followed by a rapid increase in production rate accompanying with stem-end rot development. Climacteric ethylene with a short period was associated with rapid softening and rot. Within3to5days of inoculation with B. theobromae, the decay and rot symptom completely appeared in fruit.
There are numbers of B. theobromae species that had a resistance to carbendazim. The toxicity of23fungicides to4isolates of B. theobromae was tested by mycelium growth rate methods. The toxicity results were analyzed by the EC50value, the EC90value and the cross-resistance to each other. The mango stem end rot could be effectively controlled by sporgon, propiconazole, flusilazole, prochloraz, iprodione, difenoconazole, tebuconazole, myclobutanil, pyraclostrobin and validamycin A. In addition to carbendazim, the B. theobromae also showed the resistance to other fungicides such as thiophanate-methyl, thiabendazole, kresoxim-methyl, and diniconazole. The results of cross-resistance showed that carbendazim, thiabendazole, kresoxim-methyl, and diniconazole had a positive cross-resistance for each other, while kresoxim-methyl and validamycin existed in a negative cross-resistance for each other.
Iprodione, prochloraz and iprodione.prochloraz were effective for controlling stem end rot of mango. According to overall evaluation for efficiency of controlling stem-end rot and anthracnose, iprodione.prochloraz exhibited the best capability, and the prochloraz was in the next place.
There was a significant difference (P<0.01) in toxicity of18food additives against6postharvest pathogens (C. gloeosporioidesS, C. gloeosporioidesR, B. theobromaeS, B.theobromaeR, P. mangiferae and D. dominicana) in mango fruit. The sodium o-phenylphenate was the strongest toxicity aganist6pathogens, with showing the lowest values of EC50(7.21μg/mL) and EC90value (39.53μg/mL), followed by sodium dehydroacetate and ferulic acid. The effects of sodium dehydroacetate for controlling anthracnose and stem end rot were the best among the all tested food additives, with the efficacy reaching83.52%and59.32%, respectively, followed by ferulic acid and cinnamaldehyde. The storage time of ferulic acid-treated fruit was15.51d which was the longest among the tested food additives, and followed by sodium dehydroacetate and licorice antioxidants.
The method using food additives combining with chitosan coating has been considered as a promising postharvest technology for controlling stem-end rot and anthracnose. The present results showed that the control incidences of three combination treatments including1000μ.g/mL sodium dehydroacetate+1.5%chitosan,1000μg/mL ferulic acid+1.5%chitosan and500μg/mL sodium dehydroacetate+1.5%chitosan against stem-end rot were99.34%and96.97%and90.28%respectively.
本文以可可球二孢菌(B. theobromae)等蒂腐病病菌和芒果互作为研究对象,开展芒果果实真菌潜伏侵染、B. theobromae在果实致病过程中的作用机理、B. theobromae对常用杀菌剂的抗性及其引起的采后病害防治技术等研究,旨在摸清我国芒果果实中的潜伏侵染真菌种类及严重程度,初步阐明B. theobromae致芒果果实快速腐烂的机理,探讨B. theobromae的抗药性水平及蒂腐病的安全、有效的采后处理方法,为研发芒果蒂腐病的防治措施提供依据。
本研究采用组织分离法对采自我国6省区的芒果果实潜伏侵染真菌进行检测与种类鉴定,研究结果表明：在芒果果实中普遍存在真菌的潜伏侵染现象。不同省区的带菌率存在显著差异(F=3.59,P=0.0246)。不同年份的带菌率略有差异,但未达到显著差异水平(F=1.57,P=0.2372)。果实不同部位的带菌率间存在极显著差异(F=46.44,P<0.0001),果柄和果实蒂部的带菌率极显著高于果实中部和端部的带菌率。鉴定的潜伏侵染真菌有11种,分别为Colletotrichum gloeosporioides (Penz.) Penz.&Sacc.、 Botryodiplodia theobromae Pat.、Colletotrichum acutatum Simmonds、Phomopsis mangiferae Ahmad.、Pestalotiopsis mangiferae (P. Henn.) Sutton.、Dothiorella dominicana Pet. et Cif.、 Macrophoma mangiferae Hing.、O. P. Sharma、Alternaria alternata (Fr.) Keissl.、 Aspergillus sp.、Curvularia sp.和Fusarrium spp.,其中,C. gloeosporioides和B. theobromae为芒果果实中的主要潜伏侵染真菌,M. mangiferae在芒果果实上作为潜伏侵染真菌为首次报道,B. theobromae为致病力最强的潜伏侵染真菌。
B. theobromae、C. gloeosporioides、D. dominicana和P. mangiferae4种芒果主要蒂腐病菌的致病力存在极显著差异,其中B. theobromae的致病力最强,B. theobromae在含果胶和纤维素的培养基上的生长速度均极显著高于其他3种蒂腐病菌的生长速度,采用刚果红平板水解圈法测定的结果表明,该菌产生的果胶酶活性极显著高于其他3种蒂腐病菌的果胶酶活性；纤维素酶活性虽与C. gloeosporioides无显著差异,但极显著高于D. dominicana和P. mangiferae的纤维素酶活性。采用3,5-二硝基水杨酸(DNS)比色法测定的结果表明,B. theobromae在离体培养条件下或接种与芒果果实上均可产生多聚半乳糖醛酸酶(PG)、多聚半乳糖醛酸反式消除酶(PGTE)、多聚甲基半乳糖醛酸酶(PMG)、果胶甲基反式消除酶(PMTE),以及纤维素酶(Cx)。其中PG、PMG和Cx活性较高,PGTE和PMTE活性很低。在大量产生的3种细胞壁降解酶中,其中PG活性最大,且峰值在早期出现。
B. theobromae在液体培养基中可产生导致芒果嫩叶褐变、嫩枝萎蔫和果实腐烂的致病毒素。该毒素在酸性至中性条件稳定,热稳定性强,对光照不敏感,低温条件更有利于毒素的保存。
B. theobromae在离体条件下可产生乙烯；接种到芒果果实上,在芒果果实内可诱导芒果果肉产生大量的1-氨基环丙烷-1-羧酸(ACC),从而诱导芒果果实快速释放乙烯。自然条件下发生蒂腐病的果实,在症状出现前1-2d,其乙烯释放速率即开始升高,并随着病程的发展,乙烯释放速率显著增强,且3-5d内,发病果实软化、腐烂。
B. theobromae群体中存在着对多菌灵抗性的群体。采用生长速率法进行了23种杀菌剂对4株对多菌灵抗性差异的B. theobromae的毒力测定,通过EC50值、EC90值及与多菌灵的交互抗性等的综合分析,认为咪鲜胺锰盐、咪鲜胺、丙环唑、苯醚甲环唑、氟硅唑、异菌脲、戊唑醇、腈菌唑、吡唑醚菌酯、井冈霉素,以及百菌清和代森锰锌可用作芒果蒂腐病防治的杀菌剂。B. theobromae除对多菌灵产生抗性外,还对甲基硫菌灵、噻菌灵、醚菌酯和烯唑醇4种杀菌剂产生了抗药性。通过交互抗性分析,多菌灵、噻菌灵、甲基硫菌灵和烯唑醇4种杀菌剂之间存在交互抗性,而醚菌酯与井冈霉素之间存在负交互抗性。
异菌脲、咪鲜胺和咪鲜胺·异菌脲3种杀菌剂对芒果蒂腐病具有较好的防效,而咪鲜胺·异菌脲、咪鲜胺对芒果炭疽病的防效达93%以上。根据对芒果蒂腐病和炭疽病防治效果的综合评价,咪鲜胺·异菌脲对控制芒果采后病害的效果最好,其次是咪鲜胺。
供试的18种食品添加剂对C. gloeosporioidesS、C. gloeosporioidesR、B.theobromaesS、 B. theobromaeR、P. mangiferae和D. dominicana6种芒果采后病害病原菌的毒力存在极显著差异,邻苯基苯酚钠对6种病原菌的平均ECso值和平均EC90值均最小,分别为7.21μg/mL和39.53μg/mL,其次是脱氢乙酸钠和阿魏酸。脱氢乙酸钠对炭疽病和蒂腐病的防效最好,防效分别达到83.52%和59.32%,其次阿魏酸和肉桂醛。阿魏酸处理的果实贮藏时间最长,平均贮藏时间达15.51d,其次是脱氢乙酸钠和甘草抗氧化剂,而毒力最大的邻苯基苯酚钠对芒果果实造成伤害。
使用食品添加剂结合壳聚糖的复合涂膜技术,可显著提高对蒂腐病的防治效果,1000μg/mL脱氢乙酸钠+1.5%壳聚糖涂膜与1000μg/mL阿魏酸+1.5%壳聚糖涂膜,对蒂腐病的防效分别达到99.34%和96.97%,500μg/mL脱氢乙酸钠+1.5%壳聚糖的防效达到90.28%。食品添加剂的复合涂膜技术不仅提高了对芒果蒂腐病的防治效果,还可显著降低杀菌剂的使用,且不存在农药残留问题,是一种值得推广应用的采后处理方法。
Mango (Mangifera indica Linn.), a valuable fruit crop growing throughout the tropics. However, mango is highly susceptible to postharvest decay during storage and transportation, which leads to huge economic losses. The pathogens can infect the mango flowers and immature fruit and remain quiescent until storage, during which the lesions progressively appear and symptoms become more severe along with the extension of time. It has been well known that the stem-end rot caused by Botryodiplodia theobromae is the one of most destructive postharvest disease in mango fruit.
Using B. theobromae and mango fruit as interaction system in this study, we investigated the occurrence in latent infection of pathogenic fungi in six provinces of China according to the tissue isolation method, explored the pathogenic mechanism of B. theobromae, studied the tolerance of B. theobromae to several common fungicides as well as tested some technologies to control postharvest disease in mango. The results were presented as follows:
The results showed that the latent infection of fungi comprehensively existed in the healthy mango fruits. There were significant differences in latent infection rate among the mango fruits in different provinces (F=3.59, P=0.0246), but not significant differences in latent infection rate in different years(F=1.57, P=0.2372). The rates of latent infection in the shoulder and the stem end were significantly higher than those in the other parts (F=46.44, P <0.0001). There were11pathogenic fungi to be identified, including Colletotrichum gloeosporioides (Penz.) Penz.&Sacc、Botryodiplodia theobromae Pat.、Colletotrichum acutatum Simmonds、Phomopsis mangiferae Ahmad.、Pestalotiopsis mangiferae (P. Henn.) Sutton.、Dothiorella dominicana Pet. et Cif、Macrophoma mangiferae Hing.&O. P. Sharma、 Alternaria alternata(Fr.)Keissl.、Aspergillus sp.、Curvularia sp. and Fusarrium spp.. In these isolated fungi, both C. gloeosporioides and B. theobromae were the main fungi leading to latent infection in mango. Moreover, M. mangiferae was first reported as one of latent infection fungi in mango fruit.
Four tested pathogens including B. theobromae, C. gloeosporioides, D. dominicana and P. mangiferae had different pathogenicity that cause stem end rot in mango fruit, among which the B. theobromae showed the strongest pathogenicity. The growth rate of B. theobromae in culture medium containing pectin or cellulose was extremely higher than that in other three pathogens. Furthermore, the tested results using congo red staining method showed that the activity of pectinase generated by B. theobromae was higher than that in other three pathogens, and cellulase activity produced by B. theobromae was higher than that in D. dominicana and P. mangiferae, but was similar as gloeosporioides. The results by colorimetric method of DNS demonstrated that B. theobromae may release polygalacturonase (PG), pectinmethylgalacturonase (PMG), polygalacturonic acid trans-eliminase (PGTE), pectin methyltrans-eliminase (PMTE) and cellulase (Cx.) when cultured in vitro or in vivo. Among three primary CWDEs, PG and Cx appeared activity peak earlier and activity peak of PMG was lagged.
B. theobromae in PA liquid culture produced a specific toxin that can result in browning of mango young leaves, wilting in branches and decay in fruits. The toxin had a strong thermal and acid-tolerance stability but not sensitive to light. The appropriate low temperature was favorable to toxin preservation.
B. theobromae produced ethylene in vitro, and it induced the mango fruit to produce large quantities of1-aminocyclopropane-1-carboxylic acid (ACC), which might be responsible for ethylene evolution. The ethylene began to generate increased prior to appearance in symptom of stem-end rot, followed by a rapid increase in production rate accompanying with stem-end rot development. Climacteric ethylene with a short period was associated with rapid softening and rot. Within3to5days of inoculation with B. theobromae, the decay and rot symptom completely appeared in fruit.
There are numbers of B. theobromae species that had a resistance to carbendazim. The toxicity of23fungicides to4isolates of B. theobromae was tested by mycelium growth rate methods. The toxicity results were analyzed by the EC50value, the EC90value and the cross-resistance to each other. The mango stem end rot could be effectively controlled by sporgon, propiconazole, flusilazole, prochloraz, iprodione, difenoconazole, tebuconazole, myclobutanil, pyraclostrobin and validamycin A. In addition to carbendazim, the B. theobromae also showed the resistance to other fungicides such as thiophanate-methyl, thiabendazole, kresoxim-methyl, and diniconazole. The results of cross-resistance showed that carbendazim, thiabendazole, kresoxim-methyl, and diniconazole had a positive cross-resistance for each other, while kresoxim-methyl and validamycin existed in a negative cross-resistance for each other.
Iprodione, prochloraz and iprodione.prochloraz were effective for controlling stem end rot of mango. According to overall evaluation for efficiency of controlling stem-end rot and anthracnose, iprodione.prochloraz exhibited the best capability, and the prochloraz was in the next place.
There was a significant difference (P<0.01) in toxicity of18food additives against6postharvest pathogens (C. gloeosporioidesS, C. gloeosporioidesR, B. theobromaeS, B.theobromaeR, P. mangiferae and D. dominicana) in mango fruit. The sodium o-phenylphenate was the strongest toxicity aganist6pathogens, with showing the lowest values of EC50(7.21μg/mL) and EC90value (39.53μg/mL), followed by sodium dehydroacetate and ferulic acid. The effects of sodium dehydroacetate for controlling anthracnose and stem end rot were the best among the all tested food additives, with the efficacy reaching83.52%and59.32%, respectively, followed by ferulic acid and cinnamaldehyde. The storage time of ferulic acid-treated fruit was15.51d which was the longest among the tested food additives, and followed by sodium dehydroacetate and licorice antioxidants.
The method using food additives combining with chitosan coating has been considered as a promising postharvest technology for controlling stem-end rot and anthracnose. The present results showed that the control incidences of three combination treatments including1000μ.g/mL sodium dehydroacetate+1.5%chitosan,1000μg/mL ferulic acid+1.5%chitosan and500μg/mL sodium dehydroacetate+1.5%chitosan against stem-end rot were99.34%and96.97%and90.28%respectively.
引文
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