The Establishment Of Maca Regeneration System And The Analysis Of Genetic Stability

Maca is native to Peruvian Andes （3500m above sea level） of rare medicine and food plants from high mountains. It not only contains rich protein, essential amino acids, a variety of minerals （K, Ca, Fe, zinc）, and a variety of vitamins （VB1, VB2, VC）, and other nutrients, and contains active ingredients such as Maca alkaloids, glucosinolates and sterols from secondary metabolism. A large number of studies have shown that Maca is raising fertility, improve sexual function, regulate endocrine, enhance immunity, resist oxidation, pressure resistance, fatigue resistance and anti-cancer role, and the rheumatism, respiratory disease, depression and anemia has also very good therapeutic effect. Nearly10years in Yunnan, Xinjiang, Jilin and Hunan provinces （area） Maca was introduced and cultivated successfully, and formed a certain scale of production. This has become a new growth point of forest economy in high altitude mountainous area.this paper optimize the cultivating of maca regeneration system, and ISSR molecular markers been used to tissue culture Maca genetic variation analysis, in order to provide the reference to Maca scientific production and reasonable utilization at further in our country.The main results are as follows:1、It establish maca tissue culture regeneration system（1） The germination of Maca seeds:In the5mg/L of GA3, the ratio of germination was96%after soaking12h.（2） The disinfection of Maca seeds:disinfected with0.1%HgCl₂for6min and brushed with water5times,the ratio of survival was92%.（3） Selected the best medium for maca callus induction was the MS.（4） The induction of maca Callus:used aseptic seedling leaves of maca as explant,MS＋0.1mg/L6-BA＋2.0mg/L2,4-D＋8.0g/L Agar＋25g/L Sucrose, the ratio of induction was96%.（5） The Subinoculation and multiplication of maca callus:MS+0.5mg/L NAA＋1.Omg/L6-BA+8.0g/L Ager+25g/L Sucrose, the proliferation coefficient was5.89after25d training.（6） The Differentiation of maca Callus:2.0mg/L2,4-D+O.1mg/L6-BA+2.0mg/L NAA+8.0g/LAgar＋25g/L Sucrose, the ratio of differentiation was82.67%.（7） The rooting of maca Callus:1/2MS+0.5mg/L NAA+8.0g/L Agar＋25g/L Sucrose, the ratio of rooting was88.89%.2、It built Maca plantlets ISSR-PCR amplification reactionsThis paper use orthogonal design discusses the template DNA concentration, TaqDNA polymerase concentration, Mg^2＋ concentration, dNTPs concentration, primer concentration influence on ISSR-PCR amplification reaction system,and built Maca plantlets ISSR-PCR amplification reactions:in the PCR amplification reaction system of20ul, the best condition was20ng DNA template,2.0U Taq DNA polymerase,4.0mmol/L Mg^2＋,0.10mmol/L dNTPs,0.2umol/L primer.3、The genetic stability of maca plantlets is better4、In the four subculture process of Maca plantlets,its electrophoretic bands of ISSR banding pattern consistent,qualitative change did not happen,The genetic stability of maca plantlets is better,ISSR molecular markers can be better use on studies of genetic diversity of Maca plantlets.