College of Biological Sciences and Biotechnology, Beijing Forestry University, No. 35 Tsinghua East Road, Haidian District, Beijing 100083, China.
Appl Biochem Biotechnol. 2012 Dec;168(8):2067-78. doi: 10.1007/s12010-012-9918-y. Epub 2012 Oct 18.
The inflorescences as explants for rapid propagation in vitro remained unknown in Populus euphratica Olivier. Here, we reported that multiple shoots were initiation from calli of both male and female inflorescences. The optimum medium for shoot induction from male inflorescences was lactose sulfite medium containing 1.0 mg L(-1) 6-benzylaminopurine (BA) and 0.5 mg L(-1) α-naphthalene acetic acid (NAA) or Murashige and Skoog (MS) medium containing 0.5 mg L(-1) BA and 0.2 mg L(-1) NAA. The optimum medium of shoot induction from female inflorescence calli was the MS medium containing 0.5 mg L(-1) BA and 0.2 mg L(-1) NAA. Rooting of regenerated shoots was obtained on 1/2 MS medium supplemented with 0.5∼1.0 mg L(-1) indole-3-butyric acid (IBA) and the highest frequency rooting was on medium containing 0.5 mg L(-1) IBA. No shoots were obtained on medium without BA and NAA. Peroxidase (POD) activity was measured by polyacrylamide gel electrophoresis during shoot induction and differentiation stages. The results showed that two bands of POD (2a and 2b) activity appeared lowest during the early 8 days at the dedifferentiation phase of leaves inducing calli, whereas POD 2a, 2b activity appeared to be increasing at the homeochronous dedifferentiation phase of inflorescence. Five most intensive bands, POD 1a, 1b, 1c, 2a, and ab, appeared in 8th and 28th days at the redifferentiation phase during shoot morphogenesis. These results demonstrated that the POD was involved in shoot morphogenesis from both leaf and inflorescence explants of Populus euphratica.
白杨属植物雄花序和雌花序作为外植体进行快速离体繁殖的情况尚不清楚。本文报道了从雄性和雌性花序的愈伤组织中诱导出多个芽。从雄花序诱导芽的最佳培养基是含有 1.0mg/L 6-苄基氨基嘌呤(BA)和 0.5mg/Lα-萘乙酸(NAA)的乳糖亚硫酸盐培养基或含有 0.5mg/L BA 和 0.2mg/L NAA 的 Murashige 和 Skoog(MS)培养基。从雌花序愈伤组织诱导芽的最佳培养基是含有 0.5mg/L BA 和 0.2mg/L NAA 的 MS 培养基。再生芽在添加 0.5∼1.0mg/L 吲哚丁酸(IBA)的 1/2 MS 培养基上生根,在含有 0.5mg/L IBA 的培养基上生根率最高。在不含 BA 和 NAA 的培养基上没有芽产生。在芽诱导和分化阶段通过聚丙烯酰胺凝胶电泳测量过氧化物酶(POD)活性。结果表明,在叶片诱导愈伤组织的去分化阶段的前 8 天,POD(2a 和 2b)活性出现了两条最低的带,而在花序同源去分化阶段,POD 2a、2b 活性似乎在增加。在芽形态发生的再分化阶段的第 8 天和第 28 天出现了 5 条最强烈的带,即 POD 1a、1b、1c、2a 和 ab。这些结果表明,POD 参与了白杨属植物叶片和花序外植体的芽形态发生。