| Flammulina filiformis was the fifth largest edible fungus in China.Its color was mainly yellow,light yellow or white.White F filiformis mostly introduced from Japan for industrial cultivation.Yellow F.filiformis was a traditional variety in China.Adopting farming methods.At present,yellow F.filiformis was also experimenting with industrialized cultivation,But there were fewer varieties.It was urgent to breed white F.filiformis varieties with independent intellectual property rights and yellow F.filiformis varieties suitable for industrial cultivation.In this experiment,55 germplasm resources of F.filiformis(6 yellow strains and 49 white strains)were collected at home and abroad,and comprehensive evaluation studies on somatic incompatibility,agronomic traits,esterase isoenzymes and ISSR molecular markers were carried out.Selecting the color control loci homozygous and heterozygous white and yellow F.filiformis for mon-mon hybridization,created 3 excellent germplasm.White and yellow F.filiformis with different color control loci were selected as parents,and the same nucleus,but different cytoplasmic hybrid groups were arranged,and the color of the fruiting body were investigated.The white and yellow F.filiformis homozygous for the color control loci were selected for mon-mon hybridization and backcrossing,and the fruit color of the fruiting body were investigated.The color nuclear and qualitative inheritance of F.filiformis were analyzed to improve the breeding method of F.filiformis,improve breeding efficiency,and lay a foundation for the study of color inheritance.The specific results are as follows:1.Comprehensive evaluation of screening germplasm(1)Somatic cell incompatibility(antagonistic)test showed that both yellow(colored)strains and white(colorless)strains produced antagonistic line,and yellow strains also produced antagonistic line,but only a small number of white strains were produced antagonistic line,most do not produce antagonistic line or antagonistic line was not obvious.It indicated that the genetic difference of yellow F.filiformis were large,and the genetic difference of white F.filiformis were small,and the incompatibility of somatic cells could effectively distinguish between colored and colorless F.filiformis strains.(2)A total of 15 enzyme bands with different mobility were detected by esterase isoenzyme.Cluster analysis showed that when the similarity level was 0.60,they were divided into two groups,group I were 3 strains,and group II were 52 Strain.Among them,the three strains of group I,Fv18,Fv19 and Fv20,have two specific bands(Rf 0.59 and 0.63,respectively)different from group II.The fruiting bodies were white and the base of the stalk was less adherent,and Fv18,Fv19 similarity coefficient were 1,none of the 52 strains of group II had these two specific bands.Although the color of the fruiting bodies was different(46 white strains,6 yellow strains),the base of the fruiting body had more adhesions.46 white strains can be divided into six subgroups,subgroup I was Fv7,subgroup II was Fv10,subgroup III was Fv16,subgroup IV has three strains and their bands are identical,subgroup V has 9 strains and their bands were identical,the sub-group VI has 31 strains with the same band,and the same band similarity coefficient reached 1,further indicating that the genetic difference of white F.filiformis is small,the genetic background was similar,and even other strains had the same name.And the esterase isoenzyme could not distinguish the colored and colorless F.filiformis strains.(3)The suitable temperature test for mycelial growth showed that the mycelial of the tested strains could grow in the temperature range of 5-30?,and could not grow at 35?.Among them,Fv2 has the highest growth rate at 5?,and the growth rate was also very high at 30?(the second highest).The initial screening was for the wide-temperature strain;Fv3 has the highest growth rate at 30?,and the initial screening was used to be high temperature strain.(4)Agronomic traits showed that the growth period of the tested strain was 60~121d;the color of the cap and stipe was yellow,light yellow and white,the diameter of the cap was 0.61~1.09 cm,and the diameter of stipe was 0.23~0.51 cm.The length of the stipe was 13.31~16.55 cm,the ratio of stipe and cap(the length of the stipe: the diameter of the cap)was 13.20~24.55;the average yield of the first harvest in each bag(250g dry)was 41.73~101.3g,and the yield of most white strain were higher than the yellow strain,and the highest yield was white strain Fv45,the ratio of stipe and cap was small and the numerical is 14.33,which can be used as a high-yield breeding material.(5)Twenty suitable primers were screened from 41 ISSR primers,and 267 bands were required from 55 test strains,and have 249 polymorphic bands,and the polymorphism ratio reached 93.2%.UPGMA cluster analysis showed that 55 strains were clearly divided into three groups when the similarity level was 0.78.Group I had only one wild and yellow strain Fv1,group II had only one wild and yellow strain Fv2,and group III had 53 cultivated strains.Among them,the similarity coefficient of Fv18 and Fv19 in group III reached 1,which were the same as antagonistic and esterase isoenzymes,and could be judged as the same name but different strains,but the similarity coefficient of other strains did not reach 1,indicating that ISSR can minimize the white strain which has close genetic.In summary,the genetic background of white F.filiformis was similar,and the genetic diversity of yellow F.filiformis was rich.It is difficult to distinguish the close genetic difference F.filiformis by somatic cell incompatibility and esterase isoenzyme,but ISSR can distinguish the close genetic difference between wild F.filiformis and cultivated F.filiformis.2.Innovative early maturing and high-yield germplasmWhite cultivar of F.filiformis with homozygous loci on color-controled gene and yellow cultivar with heterozygous loci were selected as parents for crossing.The monokaryotic strains with strong growth potential were selected for mon-mon hybridization.The authenticity of the monokaryotic strains and the hybrids(dikaryotic strains)were identified.Hybrids with different physiological characteristics,biochemical characteristics and agronomic traits were screened from authentic hybrids.The results showed that the isozyme bands of most hybrids were different from their parents,and the few hybrids were identical to their parents but the migration rates were different.If the similarity coefficient between the hybrids and parent strains were less 0.80,the growth period or yield of the hybrids were more likely to be transgressive inheritance.Among them,it was the yield of more white hybrids showed transgressive inheritance than their parents.The white hybrid strain HB30 was screened out with excellent commercial traits.The growth period was 6 days earlier and the yield was 4.7% higher.The excellent white hybrid strain HB95 was with good commercial traits and the yield was 15.1% high.The light yellow hybrid strain HB26 was with good commercial traits and the yield was 14.4% high.3.Color inheritance analysisFv2(homozygous yellow),Fv3(heterozygous yellow),Fv16(homozygous white)and Fv18(homozygous white)were used as parents.The nucleus was identical and cytoplasmic different by Di-mon genetic groups,and the hybrids of each hybrid group were found.The color of the fruiting body was not related to the cytoplasm and was found by Mon-mon hybridization: the hybrids of the monokaryotic strain from the yellow homozygous parent and the white monokaryotic strain were all yellow,but the color depth were different,the monokaryotic strain from the yellow heterozygous parent The color of the hybrids with the white monokaryotic strain appeared yellow(different color shade)and white,indicating that the color of F.filiformis was controlled by a single locus,but the white monokaryotic also had an effect on the color depth.A hybrid strain Fv18 and Fv2 was selected.After the cultivation,collected 50 monokaryotic strains and backcrossed with their parents.After identification,27 hybrid strains were successfully returned,and the fruiting body color were divided into yellow,light yellow and white.Further indicating that the color of Flammulina filiformis was inherited,yellow was dominant and white was recessive. |
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