红汁乳菇生物学特性与半人工栽培技术研究
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摘要
菌根性食用菌大多是自然风味鲜美独特、经济价值很高的珍稀食用菌，也是一大类难以在室内脱离共生植物完成生命周期的共生型真菌。因为其生活习性独特，栽培方式不像其它大多数腐生型食用菌那样来得便捷和易于操作。然而，其特殊的经济价值与栽培研究方面的难度也刺激和吸引国内外研究者开展多年持续不断的努力。并取得了黑孢块菌、夏块菌、红须腹菌、玉蕈离褶伞等栽培的成功。
红汁乳菇是红菇目红菇科乳菇属真菌，主要分布于中国和日本等地。是分布区内十分著名的珍贵食用菌。由于各地都有悠久的食用习惯和历史，市场交易规模较大。因此实现该菌的人工栽培是人们梦寐以求的目标。而一旦在产量上达到经济可行的水平，实现其产业化便指日可待。
本项研究即是围绕上述目标，通过野外生态因子调查，菌根、菌丝、孢子、子实体等形态显微观察、DNA中内部转录间隔区(ITS)及大亚基(LSU)序列分析的探索，较系统地研究了其生物学特性，成功栽培出了红汁乳菇子实体。设计验证了用于扩展红汁乳菇子实体和菌根等DNA的特异引物，对红汁乳菇适生土壤化学营养和物理结构进行了测定分析，为红汁乳菇种植园的建立与管理提供了依据。首次报道红汁乳菇-马尾松菌根的菌丝套网格结构特征，确认红汁乳菇菌丝细胞联接处无锁状联合；对红汁乳菇秋季产菇期间气象条件的研究表明，较大幅度的降温和降雨对诱发红汁乳菇子实体形成是必需的。特别是对红汁乳菇栽培区水分管理的研究，为保障红汁乳菇干旱年份高产稳产提供了范例。其主要研究结果如下：
1．对菌根内部结构测定显示，菌丝直径、菌丝套厚度、哈蒂氏网厚度、菌根直径的比例为1：4．7：31.9：138.4。首次报道了菌丝套的网格结构。菌丝套网格中网孔大多呈菱形，少数为不规则四边形。测量结果表明，网孔长度为宽度的2.85倍，网脊宽度为网孔宽度的16.7％。确证红汁乳菇菌丝细胞与细胞连接处没有锁状联合。
2．对多种乳菇属真菌的ITS序列和LSU序列系统发育关系的研究表明，红汁乳菇与L.akahatsu及L.sp聚成一群，但又彼此独立成枝，说明它们是亲缘关系较密切的独立的种。
Most species of the edible ectomycorrhizal fungi are precious mushrooms with unique natural zest and delicious taste and with high economical value. However, they belong to the category of co-exist fungi, which are difficult to culture without host plants at lab. Because of its special economic value and difficulty to culture, domestic and international researchers are attracted to work on it for many years. Thanks to their efforts, the cultivation of Tuber melanosprorum. Tuber aestivum. Rhizopogon rubrus, Lyophyllum and etc. are made success.
Belonging to the genus of lactarius, Russulaceae family and Russulales, Lactarius hatsutake Tanaka, a precious indigenous mushroom, is mainly distributed in China, Japan and etc., . Because of long history of using this delicious food in the distribution areas, a large scale of consuming market is formed locally. To cultivate this mushroom artificially has beenthe dream of the local people for a long time. Therefore, once its production reaches the economically acceptable level the object of industrialization will be easily realized.
The biological characters of L. hatsutake was studied systematically through investigation of ecological factors in the field, microscopic observation of its shapes of ectomycorrhizae, mycelium, spores and fruiting bodies, and analysis of its ITS and LSU segments. Based on these works, a pair of species-specific primer was designed and validated, and was proved to effective for amplifying and identifying the fructification and ectomycorrhizae of L. hatsutake. Furthermore, the characters of chemical components and physical structure of soil available for growth of L. hatsutake was surveyed and analyzed.
The major research results are showed as follows.
1. It showed by survey of structure of ectomycorrhizae that the proportion of diameter of mycelium, thickness of mantle, thickness of Hartig net, diameter of ectomycorrhizae was 1:4.7:31.9:138.4 respectively.
The shape of holes in the reticulum of mycelium mantle was almost lozenge, while a few of them were in quadrangle. The length of reticulum hole was 2.85 times of its width. And the width of reticulum ridge is equivalent to 16.7% of the width of reticulum hole.
It was ascertained that there was no clamp at the septae (connection of neighboring cells)..
2. Phylogenic relationships based on ITS and LSU sequences of species of Lactarius showed that, L. hatsutake from different areas were clustered together, and consist of a group with L.akahatsu and L.sp, but they were divided into different clades, indicated a closer consanguinity among them. However, they are, in fact, separate species.
红汁乳菇是红菇目红菇科乳菇属真菌，主要分布于中国和日本等地。是分布区内十分著名的珍贵食用菌。由于各地都有悠久的食用习惯和历史，市场交易规模较大。因此实现该菌的人工栽培是人们梦寐以求的目标。而一旦在产量上达到经济可行的水平，实现其产业化便指日可待。
本项研究即是围绕上述目标，通过野外生态因子调查，菌根、菌丝、孢子、子实体等形态显微观察、DNA中内部转录间隔区(ITS)及大亚基(LSU)序列分析的探索，较系统地研究了其生物学特性，成功栽培出了红汁乳菇子实体。设计验证了用于扩展红汁乳菇子实体和菌根等DNA的特异引物，对红汁乳菇适生土壤化学营养和物理结构进行了测定分析，为红汁乳菇种植园的建立与管理提供了依据。首次报道红汁乳菇-马尾松菌根的菌丝套网格结构特征，确认红汁乳菇菌丝细胞联接处无锁状联合；对红汁乳菇秋季产菇期间气象条件的研究表明，较大幅度的降温和降雨对诱发红汁乳菇子实体形成是必需的。特别是对红汁乳菇栽培区水分管理的研究，为保障红汁乳菇干旱年份高产稳产提供了范例。其主要研究结果如下：
1．对菌根内部结构测定显示，菌丝直径、菌丝套厚度、哈蒂氏网厚度、菌根直径的比例为1：4．7：31.9：138.4。首次报道了菌丝套的网格结构。菌丝套网格中网孔大多呈菱形，少数为不规则四边形。测量结果表明，网孔长度为宽度的2.85倍，网脊宽度为网孔宽度的16.7％。确证红汁乳菇菌丝细胞与细胞连接处没有锁状联合。
2．对多种乳菇属真菌的ITS序列和LSU序列系统发育关系的研究表明，红汁乳菇与L.akahatsu及L.sp聚成一群，但又彼此独立成枝，说明它们是亲缘关系较密切的独立的种。
Most species of the edible ectomycorrhizal fungi are precious mushrooms with unique natural zest and delicious taste and with high economical value. However, they belong to the category of co-exist fungi, which are difficult to culture without host plants at lab. Because of its special economic value and difficulty to culture, domestic and international researchers are attracted to work on it for many years. Thanks to their efforts, the cultivation of Tuber melanosprorum. Tuber aestivum. Rhizopogon rubrus, Lyophyllum and etc. are made success.
Belonging to the genus of lactarius, Russulaceae family and Russulales, Lactarius hatsutake Tanaka, a precious indigenous mushroom, is mainly distributed in China, Japan and etc., . Because of long history of using this delicious food in the distribution areas, a large scale of consuming market is formed locally. To cultivate this mushroom artificially has beenthe dream of the local people for a long time. Therefore, once its production reaches the economically acceptable level the object of industrialization will be easily realized.
The biological characters of L. hatsutake was studied systematically through investigation of ecological factors in the field, microscopic observation of its shapes of ectomycorrhizae, mycelium, spores and fruiting bodies, and analysis of its ITS and LSU segments. Based on these works, a pair of species-specific primer was designed and validated, and was proved to effective for amplifying and identifying the fructification and ectomycorrhizae of L. hatsutake. Furthermore, the characters of chemical components and physical structure of soil available for growth of L. hatsutake was surveyed and analyzed.
The major research results are showed as follows.
1. It showed by survey of structure of ectomycorrhizae that the proportion of diameter of mycelium, thickness of mantle, thickness of Hartig net, diameter of ectomycorrhizae was 1:4.7:31.9:138.4 respectively.
The shape of holes in the reticulum of mycelium mantle was almost lozenge, while a few of them were in quadrangle. The length of reticulum hole was 2.85 times of its width. And the width of reticulum ridge is equivalent to 16.7% of the width of reticulum hole.
It was ascertained that there was no clamp at the septae (connection of neighboring cells)..
2. Phylogenic relationships based on ITS and LSU sequences of species of Lactarius showed that, L. hatsutake from different areas were clustered together, and consist of a group with L.akahatsu and L.sp, but they were divided into different clades, indicated a closer consanguinity among them. However, they are, in fact, separate species.
引文
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