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作 者 ：刘艳军1；张超2；杨静慧1*；李冰2；刘婷1；秦艳筠1

期 刊 ：植物研究 2016年 36卷 1期 页码：123-128

关键词：蔗糖；培养体系；无机盐；激素；胚状体；

Keywords:sucrose, culture system, inorganic salt, hormone, embryoid,

摘 要 ：为了获得塔杨(Populus×canadensis Moench ‘Tower’)松散型、均质、胚性愈伤组织(LHEC)，以塔杨试管苗叶片为外植体，通过不同浓度激素（BA、KT、2,4-D）、蔗糖和无机盐处理，建立了松散型愈伤组织（LC）诱导—继代培养—胚性愈伤诱导的高效体系。结果显示：塔杨叶片在低蔗糖和低无机盐（1/4MS+2 mg·L^-1 2,4-D+1 mg·L^-1 BA+10g·L^-1）的培养基上培养可以诱导出完全的LC（诱导率100%）。LC在继代培养基（MS+2.0 mg·L^-1 2,4-D+2.0 mg·L^-1 BA+Vc100 mg·L^-1+30 g·L^-1）上进行2~3次转接后，再转移到LHEC培养基（MS+0.5 mg·L^-1 2,4-D+3 mg·L^-1 BA+Vc100 mg·L^-1+40 g·L^-1蔗糖）上培养4~5周(每7天转接1次)，即可诱导出具有许多球状体（原胚状体）的松散型胚性愈伤组织。此外，适当的2,4-D与BA浓度比例，可以使LC保持细胞的松散特性、不形成根、无褐变，生长快；含有30 g·L^-1蔗糖的培养基利于LC生长和LHEC的形成；维生素C能较好的抑制继代培养中的褐变；BA有利于形成LC，而KT有利于紧密型愈伤组织的形成，同时BA处理的愈伤组织生长较KT处理的快；当2,4-D浓度为0.5~1 mg·L^-1时，随着BA浓度的增加，LHEC的数量也随之增加，当BA为3 mg·L^-1时，LHEC数量达到最大值100%。本文还对影响愈伤组织胚性化的主要因素进行了讨论。提出的各项建议对均质胚性愈伤组织LHEC的培养有重要的指导意义。

Abstract:To get loose, homogeneous, embryonic callus(LHEC) from Populus×canadensis Moench ‘Tower’, an efficient system was set up from the loose callus induction(LC) to successive transfer culture and embryonic callus induction with leaves of seedlings in tube and different kinds and concentration of hormones(BA,KT,2,4-D), sucrose and inorganic salt. The loose, homogeneous callus(LC) was induced completely(induction rate of 100%) with leaves and on the culture medium of low sugar and low inorganic salt(1/4MS+2 mg·L^-1 2,4-D+1 mg·L^-1 BA+10 g·L^-1 sugar). The globule embryonic callus(formerly embryoid) was induced after LC was transferred for 2-3 times on successive culture medium(MS+2.0 mg·L^-1 2,4-D+2.0 mg·L^-1 BA+Vc 100 mg·L^-1+30 g·L^-1), then transferred onto the medium of LHEC(MS+0.5 mg·L^-1 2,4-D+3 mg·L^-1 BA+Vc 100 mg·L^-1+40 g·L^-1 sucrose), and cultivated for 4-5 weeks(one time transfer every seven days). The cells of LC was kept in the characteristics of loose without roots, browning and with fast growth under the appropriate concentration of 2,4-D and BA. LC grew better and LHEC was produced more in medium containing 30 g·L^-1 sucrose. Vitamin C suppressed browning of LC in subculture. BA was conducive to the formation of LC, but KT was beneficial to the formation of compact callus, while the callus growth was faster by treatment of BA than by KT. When the concentration of 2,4-D was 0.5-1 mg·L^-1, the number of LHEC was increased with the increase of BA concentration. The number of LHEC reached to the maximum of 100%, when BA was 3 mg·L^-1. The main factors were discussed in callus embryogenic.

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