Targeted breeding technology
Subject Category: Agronomy, Crop Breeding and Breeding, code 210.3035
The breeding of edible fungi is achieved by means of spore hybridization, protoplast fusion of remote strains and gene improvement. The new varieties of polygenic polymerization are selected with the help of molecular markers and enzymatic characteristics, and the excellent strains are created.
Advantages:
1) The maximum of experimental varieties on the base: the huge number of own bases + agent bases makes the progress of pilot, pilot, optimization and promotion of varieties very short, and the collection and analysis of ownership process data can be successfully completed within one year, and the process data information management system established accordingly can be effectively used in product after-sales service.
2) Regional selection for directional breeding: Taking Liaoning region as the test sampling point, covering the feedbacks of breeding data from agent bases in eastern Liaoning, southern Liaoning and western Liaoning, screening strains suitable for cultivation in different regions, and realizing directional breeding according to local conditions.
Technical route:
The technical team selected two methods: monospore cross breeding with strong breeding direction and protoplast fusion breeding with greater potential to produce new strains:
1) Single spore cross breeding
The basic principle of hybrid breeding of edible fungi is the gene recombination in the process of cell meiosis. This breeding method is suitable for edible fungi with heterogeneic combination, such as oyster mushroom, shiitake mushroom, fungus, monkey head, enoki mushroom and so on. The strain formed by spore germination of heterozygous edible fungi did not have fruiting ability, and the fruiting body could not be formed without hybridization with the friendly spore strains. Only through the combination of different mononuclear mycelium pairs, can the dinucleation form the fruiting body. According to this principle, using mononuclear mycelium hybridization with different genetic advantages to breed excellent hybrid heterokaryons is an important way of edible fungi breeding, which has stronger direction and operability than mutagenesis breeding.
The operation steps of single spore hybridization: single spore acquisition → mononuclear mycelium germination → mononuclear mycelium hybridization affinity → selection of hybrid binary mycelium → parent antagonism verification → hybrid strain fruiting verification → character evaluation
2) protoplast fusion breeding
Protoplast fusion is the exchange and recombination of the whole genome through the cell fusion induced by the fusion agent of the protoplasts of different genetic types after the removal of the cell wall to produce new varieties and types. Protoplast fusion can be carried out between species of the same family. Compared with single spore hybridization breeding, the interspecific span is larger, but the stability of the fusion is less than that of the heterozygote formed by single spore hybridization.
Protoplast fusion procedure: protoplasmic preparation → genetic labeling → protoplast fusion → medium regeneration → parent antagonism test → fruiting verification → trait evaluation → genetic stability
Comparison test of three-stage efficient varieties of edible fungi:
1) Take a small test
The new strains were selected for the same shed ratio test. Each shed cultivated at most 4 varieties at the same time, placed at intervals, and 3 parallel repeats were performed (ABCD; ABCD; ABCD), make a comprehensive evaluation of the agronomic traits of each variety, and select the dominant varieties for pilot cultivation.
2) Pilot test
The medium scale fruiting of the varieties to be tested was verified, and one variety was cultivated independently in each shed to evaluate the agronomic and commercial traits of the fruiting bodies. After the fruiting body was harvested, the seed strains were obtained by tissue separation method, and the mushrooms were tested by the same shed. The agronomic and commercial traits of the fruiting body were evaluated, and the genetic stability of the strains was tested.
3) Batch test
The test varieties were tested in batches, and 1 variety was cultivated independently in each shed, and at least 10 sheds were tested in batches to evaluate the agronomic and commercial traits of fruiting bodies. Investigate whether the varieties to be tested have special requirements for facilities, equipment, manpower and other resources, investigate market feedback information, and determine whether the varieties are suitable for promotion and application, and whether there is variety certification and protection value.