基于葡萄柚葉片離體再生體系的研究

陶燕藍　藍增全　吳田

摘 要 以10個葡萄柚品種種子萌發的無菌苗葉片為外植體，研究葡萄柚的離體再生體系。結果表明：在MS+1.0 mg/L 2，4-D+6%蔗糖培養基上，8個葡萄柚品種能誘導出愈傷組織，其中“星路比”品種誘導率最高為46.67%，將誘導的愈傷組織轉移至相同培養基中可繼代增殖；“帕利斯”、“星路比”、“泰國種”3個品種的葉愈傷組織接種于MS+3.0 mg/L 6-BA分化培養基上可分化出叢生芽，這些叢生芽在MS+0.2 mg/L 6-BA培養基上平均可增殖4～6個芽；叢生芽在1/2 MS+1.0 mg/L IBA生根培養基上可誘導出不定根。煉苗后，田間移栽成活率為78.18%。為后續葡萄柚良種選育及遺傳轉化體系的建立奠定基礎。

關鍵詞 葡萄柚；再生植株；愈傷組織；不定芽

中圖分類號 Q943.1 文獻標識碼 A

Abstract The leaves of aseptic seedlings from the seeds of ten grapefruit varieties as explants were used to establish the in vitro regeneration system of grapefruit. The results showed that the callus of eight grapefruit varieties were observed in the medium MS+1.0 mg/L 2，4-D+6% sugar， and “Star Ruby”had the highest callus induction frequency， which is 46.67%. The multiplication medium of the callus was the same medium. The medium of the differentiation of adventitious buds of three grapefruit varieties“Perlist”“Star Ruby”and “Thailand” was MS+3.0 mg/L 6-BA. The medium for multiplication of the regeneration buds was MS+0.2 mg/L 6-BA， and the rooting medium was 1/2 MS+1.0 mg/L IBA. The survival rate of regenerative plant was 78.18%. In order to provide useful references for the breeding and the application of genetic transformation technique in grapefruit.

Key words Citrus paradisi Macf.；Regenerative plant；Callus；Adventitious bud

doi 10.3969/j.issn.1000-2561.2015.03.007

葡萄柚（Citrus paradisi Macf.）是蕓香科柑橘屬亞熱帶常綠果樹，為甜橙和柚的天然雜交種，是世界柑桔類四大類群之一[1]，因果實懸掛成串，簇生如葡萄，故稱葡萄柚[2]。葡萄柚起源于西印度群島的巴巴多斯，在歐美地區大量種植，中國種植較晚[3]。葡萄柚的營養價值豐富、果實柔嫩、風味獨特，既可鮮食，又可加工制成罐頭、果汁及色拉等原料[4-6]，越來越受到世界各國的生產者和消費者的喜愛，因此發展葡萄柚種植具有可觀的前景[7]。

葡萄柚主要通過接穗嫁接栽培[8]，不僅費工費時，而且易造成品種退化，給葡萄柚的生產帶來損失。通過建立高效的離體再生體系，能有效克服傳統繁殖方式的弊端，為葡萄柚的原體苗木遺傳育種、基因轉化及品種改良提供重要基礎。葉片是植物光合作用的重……