Pollen Culture Of The Three Alfalfa Varieties And Its Haploid Identification

The obtaining of haploid through alfalfa anther culture or pollen culture and the further hybrid breeding with pure materials can fully play the advantage of the hybrid,shorten the breeding period and improve breeding efficiency.In this study,the tissue culture of the anther of Medicago varia Martin.cv.Caoyuan No.1,Medicago falcata L.cv.Hulunbeier and Medicago sativa L.cv.Xinjiang Daye was carried out,and the induction of anther callus was conducted through orthogonal test by employing solid culture and liquid suspension culture media in different hormone combination;the regenerated plants were obtained after differentiation and rooting culture,and appropriate anther tissue regeneration system was established;the ploidy of the regenerated plants were determined by the flow cytometry identification of pore number primary selection and chromosomes at root tip.The main results are as follows:(1)In the induction of anther callus,solid medium was superior to liquid suspension medium for Medicago varia Martin.cv.Caoyuan No.1,and liquid suspension medium was superior to solid medium for Medicago falcata L.cv.Hulunbeier and Medicago sativa L.cv.Xinjiang Daye,the highest callus induction rate value were 83.33%,70.00%and 83.33%.According to the range analysis of orthogonal test.The medium suitable for Medicago varia Martin.cv.Caoyuan No.1 was B5 + 0.4 mg/L NAA+ 0.75 mg/L 2,4-D + 1.0 mg/L KT + 0.25 mg/L 6-BA;the medium suitable for Medicago falcata L.cv.Hulunbeier is B5 + 0.4 mg/L NAA + 0.50 mg/L 2,4-D + 3.0 mg/L KT + 0.25 mg/L 6-BA;the medium suitable for Medicago sativa L.cv.Xinjiang Daye was MS + 0.2 mg/L NAA+ 2.0 mg/L 2,4-D + 3.0 mg/LKT+ 0.5mg/L 6-BA.(2)The main hormones affecting the callus formation of the three types of alfalfa were different.In the process of solid culture,the hormone of Medicago varia Martin.cv.Caoyuan No.1 was KT,and the hormones of Medicago falcata L.cv.Hulunbeier and Medicago sativa L.cv.Xinjiang Daye were 2,4-D;in the process of liquid suspension culture,the hormone of the sterile plants of Medicago varia Martin.cv.Caoyuan No.1 was KT and that of the fertile plants was 6-BA,and the hormones of Medicago falcata cv.Hulunbeier and Medicago saliva L.cv.Xinjiang Daye were 2,4-D.(3)The medium suitable for the callus differentiation culture of the three types of alfalfa was MS + 1.0 mg/L KT + 0.2 mg/L NAA,and the differentiation rate is 58.33%-73.33%.(4)The numbers of haploid plants obtained from the anther culture regenerated plants of sterile plants of Medicago varia Martin,cv.Caoyuan No.1,Medicago falcata L.cv.Hulunbeier and Medicago sativa L.cv.Xinjiang Daye were four,one and seven,accounting for 8.5 1%,20%and 14%of the total number.