Isolation and identification of Lactarius deliciosus and the interaction with Pinus massoniana

【Objective】To conduct isolation and identification of Lactarius deliciosus and cocultivation of L. deliciosus and Pinus massoniana, so as to realize large-scale cultivation of L. deliciosus by the co-cultivation.【Method】The fruiting body of Lactarius matsutake was collected from the natural masson pine secondary forest in Enshi, Hubei Province, and the Lactarius matsutake strain was obtained by tissue separation, genus and species of the strain were isolated through rDNA ITS. Influences of L. deliciosus inoculation on growth, mycorrhizal infection rate, nitrogen(N), phosphorus(P) and Kalium(K) contents of P. massoniana were analyzed using one-way ANOVA, and correlation analysis was conducted using Duncan method to explore correlation between the mycorrhizal infection rate of P. massoniana and the growth of L. deliciosus.【Result】According to the results of molecular identification of rDNA ITS sequences, three stains of fungi were identified as L. deliciosus. The L. deliciosus strains grew well on media such as pine sawdust and cottonseed shells. L. deliciosus strains were partially saprobe. Co-cultivation of L. deliciosus and P. massoniana showed that 3 L. deliciosus strains had developed mycorrhizae with P. massoniana. After co-cultivation for 11 months, the mycorrhizal infection rate of P. massoniana with L. deliciosus inoculation were all above 95%. Dry weight, height, basal diameter, length of main root and number of lateral roots were significantly increased(P<0.05, the same below). Compared with CK, the dry weights, plant heights, basal diameters, lengths of main roots and numbers of lateral roots of P. massoniana in treatments were increased by 5.41%-19.25%, 11.15%-20.59%, 12.58%-28.90%, 34.88%-46.42% and 52.63%-68.42%, respectively. Contents of N and P elements of P. massoniana were promoted with exogenous inoculation of L. deliciosus, which was good for cultivation of P. massoniana seedling growth. Generally, inoculation with L-1 had better effect on mycorrhizal infection rate, growth and absorption of N and P in P. massoniana than L-3 and L-2. After co-cultivation for 13 months, twenty-four fruiting bodies were obtained, with an average output of 2.67 mushrooms/m² and a yield of 80.67 g/m². However, the correlations of average number and total weight of fruiting bodies of L. deliciosus with the mycorrhizal ratio of P. massoniana were not significant(P>0.05).【Conclusion】rDNA ITS can be used to identify wild L. deliciosus accurately. The growth of P. massoniana and utilization of N and P were promoted by L. deliciosus exogenous inoculation. At the same time, the increase of mycorrhizal infection rate is helpful for the formation of fruiting body of L. deliciosus. The L. deliciosus in Enshi, Hubei Province can be used as a potential species for cultivation of L. deliciosus.