红肉猕猴桃再生体系的建立及DNA条形码鉴定

Establishment of regeneration system and DNA barcode identification in red-fleshed kiwifruit

以产于河南省栾川县的野生红肉猕猴桃(Actinidia sp.)叶片为外植体,诱导产生愈伤组织,并对其进行不定芽分化、不定芽增殖、无菌苗生根等方面的研究,建立了该品种再生体系;同时运用DNA条形码技术,以ITS、mat K、SQ1、ML为目标序列,对其亲缘关系进行鉴定,结果显示:在MS+0.5 mg·L^-1 6-苄氨基嘌呤(6-BA)+2.0 mg·L^-1 2,4-二氯苯氧乙酸(2,4-D)培养基中,愈伤组织分化率达100%;在MS+4 mg·L^-1 6-BA+0.2 mg·L^-1萘乙酸(NAA)培养基中, 49 d后,不定芽分化率达88.89%;在MS+3.5 mg·L^-1 6-BA+0.2 mg·L^-1 NAA+0.2 mg·L^-1赤霉素(GA3)中,不定芽增殖率达100%,增殖系数为7.85;在1/2MS+0.2 mg·L^-1吲哚丁酸(IBA)培养基中,不定芽生根率达到92.59%,且根系发达;移栽到珍珠岩与田间土壤体积比为1:4的生根基质中, 14 d后移栽成活率达到94%;DNA条形码分析结果显示产于河南省栾川县的红肉猕猴桃属于猕猴桃属净果组片髓系的软枣猕猴桃(A. arguta)。

An in vitro propagation system was developed from leaf explants of red-fleshed kiwifruit(Actinidia sp.) from Luanchuan County in Henan Province on the basis of the callus and shoot differentiation, shoot multiplication and rooting culture. Furthermore, molecular biological identification for genetic relationship of redfleshed kiwifruit with its relative species was carried out by four DNA barcodes(ITS, matK, SQ1 and ML). The results show that the rate of callus differentiation of the leaf explants reached to 100% on MS+0.5 mg·L^-1 6-BA +2.0 mg·L^-1 2,4-D. The rate of adventitious bud differentiation was 88.89% on MS+4 mg·L^-1 6-BA+0.2 mg·L^-1 NAA in a 49-day culture period. Cultured on MS+3.5 mg·L^-1 6-BA+0.2 mg·L^-1 NAA+0.2 mg·L^-1 GA3, the proliferation rate of adventitious buds and the proliferation coefficient were 100% and 7.85, respectively. The rooting rate of adventitious buds was 92.59% with root flourishing on 1/2 MS+0.2 mg·L^-1 IBA. The survival rate of transplantation was 94% in the mixed matrix of pearlite and soil(1:4, V/V) in a 14-day culture period. The redfleshed kiwifruit from Luanchuan County in Henan Province belongs to A. arguta(Ser. Lamellatae, Sect. Leiocarpae).