Construction Of Sugarcane Mutant Library Induced By EMS And Screening Of SSR Primers

In this study,Hunan hanshou native sugarcane varieties were used as experimental materials,constructing its tissue culture system,the effects of different concentration of EMS solution on the mutagenesis of callus,tissue cultured buds and field shoot were studied,screening the optimal concentration of mutagenesis.At the same time,the mutant plants can be selected accurately,SSR primers were screened with 95 sugarcane materials.Finally,the selected SSR core primers were used to identify the mutagenic materials.At the same time,the field agronomic characters of the mutant were evaluated.The main results are as follows:1.In the construction of sugarcane tissue culture system,different concentrations of 2,4-D on the callus induction rate of high and low order is3.0mg/L>3.5mg/L>4.0mg/L>2.5mg/L>2.0mg/L,the maximum differentiation rate was 93.92%;When 6-BA and NAA were added to callus differentiation medium,the average differentiation rate of sugarcane callus was between 70.00% and 95.00%,When NAA and KT were added,the average differentiation rate was lower than 70.00%,6-BA was more effective than KT;6-BA,NAA and KT hormone combinations with different concentrations are added into the culture medium for sterile bud proliferation,in a certain range,the higher the concentration,the greater the proliferation coefficient,but it will decrease after reaching a certain value.2.When EMS mutagenize sugarcane callus,tissue culture bud and field stem bud,in the case of processing the same time,with the increase of the solubility of EMS solution,the growth potential showed a downward trend;Similarly,when the concentration of EMS mutation is the same,the growth potential also shows a downward trend with the increase of treatment time.3.From 68 pairs of SSR primers published,21 pairs of SSR primers with high polymorphism.The PIC value of 21 pairs of primers was 0.9279,the minimum value was 0.7855,and the average value was0.8664.In this study,the polymorphism of 21 pairs of core primers was verified with 95 materials from19 populations.The results showed that 19 populations could be divided into 4 subgroups by UPGMA cluster map,at the same time,the population structure was divided into 6 subgroups,95 validation materials showed rich diversity in 21 pairs of primers.21 pairs of SSR primers can be used to distinguish the materials of this study,and can be used for the detection of mutant materials.4.A total of 320 plantlets from EMS-induced calli and buds survived were amplified by 21 pairs of primers,not genetic variation plants were detected.While for the field shoots treated with EMS solution,a total of 1000 plants were screened for polymorphism amplification,and 31 mutants were detected.5.Seven agronomic traits of mutant and control plants were investigated,and grey correlation analysis was carried out.The results showed that the correlation between 31 mutants and control materials was greater than 0.7.The mutant material with the highest correlation degree was Y808,and the mutant material with the lowest correlation degree was Y162.