Sekeli Rogayah, Abdullah Janna Ong, Namasivayam Parameswari, Muda Pauziah, Abu Bakar Umi Kalsom
Department of Microbiology, Faculty of Biotechnology & Biomolecular Sciences, Universiti Putra Malaysia, Selangor Darul Ehsan, 43400 UPM Serdang, Malaysia.
Department of Cell & Molecular Biology, Faculty of Biotechnology & Biomolecular Sciences, Universiti Putra Malaysia, Selangor Darul Ehsan, 43400 UPM Serdang, Malaysia.
ISRN Biotechnol. 2012 Oct 21;2013:958945. doi: 10.5402/2013/958945. eCollection 2013.
A high survival rate for transformed papaya plants when transferred to the field is useful in the quest for improving the commercial quality traits. We report in this paper an improved rooting method for the production of transformed Malaysian Eksotika papaya with high survival rate when transferred to the field. Shoots were regenerated from embryogenic calli transformed with antisense and RNAi constructs of 1-aminocyclopropane-1-carboxylic acid oxidase (ACO) genes using the Agrobacterium tumefaciens-mediated transformation method. Regenerated transformed shoots, each measuring approximately 3-4 cm in height, were cultured in liquid half-strength Murashige and Skoog (MS) medium or sterile distilled water, and with either perlite or vermiculite supplementation. All the culturing processes were conducted either under sterile or nonsterile condition. The results showed that rooting under sterile condition was better. Shoots cultured in half-strength MS medium supplemented with vermiculite exhibited a 92.5% rooting efficiency while perlite showed 77.5%. The survival rate of the vermiculite-grown transformed papaya plantlets after transfer into soil, contained in polybags, was 94%, and the rate after transfer into the ground was 92%. Morpho-histological analyses revealed that the tap roots were more compact, which might have contributed to the high survival rates of the plantlets.
转化后的木瓜植株移栽到田间后具有较高的成活率,这对于改善商业品质性状很有帮助。我们在本文中报道了一种改进的生根方法,用于培育转化后的马来西亚埃克索蒂卡木瓜,使其移栽到田间后具有较高的成活率。使用根癌农杆菌介导的转化方法,从用1-氨基环丙烷-1-羧酸氧化酶(ACO)基因的反义构建体和RNAi构建体转化的胚性愈伤组织中再生芽。将再生的转化芽(每个芽高约3-4厘米)在液体半强度Murashige和Skoog(MS)培养基或无菌蒸馏水中培养,并添加珍珠岩或蛭石。所有培养过程均在无菌或非无菌条件下进行。结果表明,在无菌条件下生根效果更好。在添加蛭石的半强度MS培养基中培养的芽生根效率为92.5%,而添加珍珠岩的为77.5%。装在塑料袋中的蛭石培育的转化木瓜幼苗移栽到土壤后的成活率为94%,移栽到地里后的成活率为92%。形态组织学分析表明,主根更加紧密,这可能是幼苗成活率高的原因。