摘要
金花茶(Camellia nitidissima)为山茶科山茶属植物,属于国家一级保护植物,具有很高的观赏价值及药用价值。但目前金花茶野生资源少,因此寻找高效的人工种植方法快速繁殖金花茶优良种苗,进而大幅度提高规模化种植金花茶的产量十分必要。丛枝菌根(arbuscular mycorrhiza,AM)真菌能显著促进植物生长发育、提高作物产量、增强植物抗逆性和抗病性,金花茶接种AM真菌,对金花茶有积极的影响。由于几乎所有的AM真菌接种试验均涉及菌根发育状况观察与侵染率测定,而目前AM侵染状况的观察测定方法种类繁多,良莠不齐,因此本研究探究一种金花茶根系AM真菌检测的最佳染色方法,以便更好地观察AM真菌对金花茶根系的侵染情况。本研究以金花茶健康植株根系为试验材料,探讨5种染色剂(苏丹红Ⅳ、酸性品红、苯胺蓝、台酚蓝、醋酸墨水)对金花茶根系AM真菌的染色效果。研究表明:5%醋酸墨水染色液的染色效果最佳,根皮层细胞内AM真菌的菌丝、泡囊、孢子等结构清晰可见,且能够明确分辨AM真菌与其他未知真菌,根的染色效果可以长久保存;利用苏丹红Ⅳ和酸性品红染色的AM真菌菌丝、泡囊与背景反差不明显,而且酸性品红的染色效果保存时间短,不利于观察;苯胺蓝和台酚蓝的染色效果略次于醋酸墨水,且价格昂贵,成本高。除醋酸墨水外,其他4种染色剂均是疑似致癌物,长期使用对操作人员的健康产生非常大的安全隐患。醋酸墨水染色操作简便、毒性低、成本低廉、染色效果极佳,适用于金花茶根系AM真菌的染色和制片观察。本研究结果可为今后金花茶菌根种苗的应用提供新的思路。
Camellia nitidissima belongs to the genus Camellia of theaceae and belongs to the first-level national protection plant,with high ornamental value and medicinal value.However,at present,there are few wild resources of C.nitidissima.It is necessary to find efficient artificial cultivation methods,rapidly propagate excellent C.nitidissima seedlings,and then greatly improve the yield of large-scale cultivation of C.nitidissima.Arbuscular mycorrhizal(AM)fungi can significantly promote plant growth and development,increase crop yield,and enhance plant stress and disease resistance.Inoculation of AM fungi into C.nitidissima has a positive effect on the cultivation of C.nitidissima.Since almost all AM fungal inoculation test involves the growth of mycorrhizal observation and infection rate determination,and AM infection situation observation methods available are not efficient.An efficient dyeing of AM fungi C.nitidissima was reported.In this study,the effects of five dyes(Sudan redⅣ,acid fuchsin,trypan blue,aniline blue and acetic acid ink)on AM fungi of healthy plant roots of C.nitidissima were investigated.The results showed that 5%acetic acid ink staining solution had the best staining effect.Mycelia,vesicles,spores and other structures of AM fungi in root cortex cells were clearly visible,and AM fungi could be clearly distinguished from other unknown fungi,and the root staining effect could be preserved for a long time.The mycelia and vesicles of AM fungi stained with Sudan redⅣand acid fuchsin were not in obvious contrast with the background,and the preservation time of acid fuchsin staining was short,which was not conducive to observation.The dyeing effect of aniline blue and trypan blue was slightly inferior to acetic acid ink,and the price was expensive and the experimental cost was high.The last four dyes are suspected of causing cancer,and the long-term use has great potential safety risks to the health of operators.Acetic acid ink dyeing is simple,low toxicity,low cost and excellent dyeing effect.It is suitable for the staining and section observation of AM fungi in the roots of C.nitidissima.The results would provide new ideas for the future application of C.nitidissima mycorrhizal seedlings.
作者
许诗萍
高日芳
张金莲
宋娟
李冬萍
何苑皞
曾诗媛
陈廷速
XU Shiping;GAO Rifang;ZHANG Jinlian;SONG Juan;LI Dongping;HE Yuanhao;ZENG Shiyuan;CHEN Tingsu(College of Forestry,Central South University of Forestry and Technology,Changsha,Hunan 410004,China;Institute of Micro-biology,Guangxi Academy of Agricultural Science,Nanning,Guangxi 530007,China;Yulin Normal University,Yulin,Guangxi 537000,China)
出处
《热带作物学报》
CSCD
北大核心
2024年第1期215-224,共10页
Chinese Journal of Tropical Crops
基金
广西科技基地和人才专项(桂科AD20159001)
广西农业科学院基本科研业务专项(桂农科2021YT097)。
关键词
金花茶
根系
丛枝菌根真菌
染色
Camellia nitidissima
roots
arbuscular mycorrhiza(AM)fungi
staining
