Tang Wei, Newton Ronald J
Lab of Conifer Biotechnology, Department of Biology, Howell Science Complex, East Carolina University, Greenville, NC 27858-4353, USA.
Plant Physiol Biochem. 2005 Aug;43(8):760-9. doi: 10.1016/j.plaphy.2005.05.008. Epub 2005 Jun 13.
We reported establishment of an efficient plant regeneration procedure through direct adventitious shoot (DAS) formation from cotyledons and hypocotyls of eastern white pine (Pinus strobus L.) mature embryos in this investigation. Multiple DASs were initiated from cotyledons of embryos on PS medium containing N6-benzyladenine (BA), thidiazuron (TDZ), or kinetin (KIN). Among different concentrations of casein enzymatic hydrosylate (CH) and glutamine used in this study, 500 mg l(-1) CH or 600 mg l(-1) glutamine induced the highest frequency of DAS formation. Rooting of regenerated shoots was obtained on PS medium supplemented with 0.01-0.1 microM indole-3-acetic acid (IAA) with the highest frequency on medium containing 0.01 muM IAA. No DASs were obtained on medium without TDZ. Measurement of peroxidase (POD) and catalase (CAT) activity during direct shoot induction and differentiation demonstrated that the lowest POD activity appeared in the 5-6th week of culture and lowest CAT activity occurred in the 7-8th week of culture on medium with TDZ. No such a change in POD and CAT activities was observed on medium without TDZ. These results demonstrated that POD and CAT activities were involved in DAS formation induced by TDZ in eastern white pine.
在本研究中,我们报道了通过东部白松(Pinus strobus L.)成熟胚子叶和下胚轴直接不定芽(DAS)形成建立高效植物再生程序的方法。在含有N6-苄基腺嘌呤(BA)、噻二唑素(TDZ)或激动素(KIN)的PS培养基上,从胚子叶诱导出多个DAS。在本研究使用的不同浓度的酪蛋白水解物(CH)和谷氨酰胺中,500 mg l(-1) CH或600 mg l(-1)谷氨酰胺诱导的DAS形成频率最高。在添加0.01 - 0.1 microM吲哚-3-乙酸(IAA)的PS培养基上可使再生芽生根,在含有0.01 muM IAA的培养基上生根频率最高。在不含TDZ的培养基上未获得DAS。在直接芽诱导和分化过程中对过氧化物酶(POD)和过氧化氢酶(CAT)活性的测定表明,在含有TDZ的培养基上,培养第5 - 6周时POD活性最低,培养第7 - 8周时CAT活性最低。在不含TDZ的培养基上未观察到POD和CAT活性的这种变化。这些结果表明,POD和CAT活性参与了TDZ诱导的东部白松DAS形成。