| Arbuscular mycorrhizal(AM)fungi can form a mutually beneficial symbiosis with more than 80% of terrestrial vascular plants on the earth,which can promote plants to absorb soil mineral nutrients,improve plant resistance and disease resistance,improve soil structure,repair soil heavy metal pollution,etc.,and can be used as natural Bacterial fertilizer can also be used as a biocontrol agent,and it has great application prospects in agricultural(forest)production.However,the obligatory symbiosis of AM fungi makes it necessary to grow and complete the life history in symbiosis with the host plant root system,which greatly limits the large-scale production of AM fungal inoculants.Using hairy roots as the host,inoculating AM fungal spores,and establishing a hairy root-AM fungus dual culture system is an effective way to solve the problem of AM fungus pure culture and large-scale production of inoculants.In this experiment,Agrobacterium rhizogenes C58C1 induced 4 kinds of tobacco and belladonna to produce hairy roots and Septoglomus spores isolated and identified from the soil to establish a dual culture system,and selected suitable Septoglomus spores According to the disinfection method and culture temperature,three kinds of tobacco hairy roots and Septoglomus dual culture system were successfully established,and the optimal host hairy roots of Septoglomus were screened out.The specific test results are as follows:1.The leaves of NC82,VA116,K87,K326 and Belladonna were infected by C58C1,which were identified as hairy roots by morphological and histological examination.The hairy root induction rate(K87(92.86%)> K326(89.61%)>NC82(87.34%)> Belladonna(78.46%)> VA116(38.64%)was different among the five plant genotypes.It was concluded that reducing the sucrose concentration was more beneficial to the growth of hairy roots.2.The spores were isolated from the soil and identified by morphology to be Se.constrictum and Se.furcatum through morphological identification.Among them,Se.furcatum is a new record species in China.It was found that 75% alcohol could effectively reduce the contamination rate.Combined with traditional disinfection method,short time(0.5 ? 1 min)75% alcohol and low temperature treatment were more favorable to spore germination.The treatments with the highest germination rate(48.06%)were: 75% alcohol for 30 seconds,a solution and B solution for 6 minutes,4 ? for one week and then transferred to 25 ?.When spores germinate,hyphae continue to grow from the hyphae,not far from the spores hyphae will produce a large number of branches,easy to form near the spores hyphae cluster.3.Different plant species and different plant genotypes have different symbiosis between hairy roots and spores.The results showed that the hairy roots of NC82,Va116,K87 and Septoglomus had established a double culture system,no spores were produced,no symbiosis existed between VA116 and Belladonna.The NC82 hairy root is the best host.In the course of spore development,it was found that the hyphae end of hyphae expanded in the absence of symbiosis,which was similar to the sporulation of other species of AM fungi,and in the course of hyphae growth,specific structures such as healing,annular hyphae and triangular cross hyphae were observed.In this experiment,five stable hairy root induction systems were established,and a large number of stable hairy roots were obtained,which provided a good host material for subsequent research;the first study Septoglomus spore disinfection method and germination temperature;The dual culture system was established with belladonna hairy root as the host for the first time,but was unsuccessful;the tobacco NC82,K87,Va116 three kinds of tobacco hairy roots and Septoglomus dual culture were successfully established,but no spores were produced.The spore development and mycelial growth process were recorded,and the tobacco NC82 hairy root was determined to be the most suitable host for Septoglomus. |
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