Effects of plant growth regulators, carbon sources and pH values on callus induction in Aquilaria malaccensis leaf explants and characteristics of the resultant calli
Effects of plant growth regulators, carbon sources and pH values on callus induction in Aquilaria malaccensis leaf explants and characteristics of the resultant calli
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DOI：
10.1007/s11676-014-0492-8
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发表时间：
2014-09-01
影响因子：
3
通讯作者：
Mohamed, Rozi
中科院分区：
文献类型：
--
作者：
Jayaraman, Shashita;Daud, Nurul Hazwani;Mohamed, Rozi
濒临灭绝的热带树木，Aquilaria malaccensis，生产。沉香木用于香料和药物。目前正在努力通过组织培养生产有价值的沉香化合物。本研究的目的是开发一种最佳的生长培养基，特别是最佳激素组合的愈伤组织悬浮培养。利用苗圃栽培的A. Malaccensis，首先将灭菌的叶外植体在含有15 g/L蔗糖和pH 5.7的基本Murashige和Skoog（MS）凝胶培养基上孵育。不同的生长素类型，包括1-萘乙酸（NAA），2，4-二氯苯氧乙酸（2，4-D），吲哚-3-丁酸（IBA），进行了测试，在不同的浓度（0.55，1.1和1.65 μ M）使用的基本培养基。将叶外植体在黑暗中孵育30天。1.1 μ M NAA诱导的愈伤组织生物量最高，达17.3mg，但愈伤组织为致密型。然后将该生长素浓度与0.55、1.1、2.2或3.3 μ M的6-苄氨基嘌呤（BAP）或激动素组合以诱导易碎愈伤组织的生长。1.1 μ M NAA + 2.2 μ M BAP组合产生的愈伤组织较疏松，生物量最高（93.3mgDW）。当测试不同碳源和pH值时，蔗糖15 g/L和pH值5.7产生最高生物量，分别为87.7mg和83 mg DW。显微镜下观察显示，松散的细胞排列为相对较大的细胞，而对于致密的愈伤组织，细胞较小且密集排列。结果表明，MS+蔗糖15 g/L，NAA 1.1 μ M + BAP 2.2 μ M，pH值5.7的培养基是诱导紫云英叶片愈伤组织的最佳培养基。Malaccensis的细胞悬浮培养。
The endangered tropical tree, Aquilaria malaccensis, produces. agarwood for use in fragrance and medicines. Efforts are currently underway to produce valuable agarwood compoundsn tissue culture. The purpose of this study was to develop an optimal growth medium, specifically, the best hormone combination for callus suspension culture. Using nursery-grown A. malaccensis, sterilized leaf explants were first incubated on basic Murashige and Skoog (MS) gel medium containing 15g/L sucrose and at pH 5.7. Different auxin types including 1-naphthaleneacetic acid (NAA), 2,4-dichlorophenoxyacetic acid (2,4-D), and indole-3-butyric acid (IBA), were tested at various concentrations (0.55, 1.1 and 1.65 mu M) using the basic medium. Leaf explants were incubated for 30 days in the dark. Callus induced by 1.1 mu M NAA had the highest biomass dry weight (DW) of 17.3 mg; however the callus was of a compact type. This auxin concentration was then combined with either 6-benzylaminopurine (BAP) or kinetin at 0.55, 1.1, 2.2 or 3.3 mu M to induce growth of friable callus. The 1.1 mu M NAA + 2.2 mu M BAP combination produced friable callus with the highest biomass (93.3mg DW). When testing the different carbon sources and pHs, sucrose at 15g/L and pH at 5.7 yielded highest biomasses at 87.7mg and 83mg DW, respectively. Microscopic observations revealed the arrangement of the friable cells as loosely packed with relatively large cells, while for the compact callus, the cells were small and densely packed. We concluded that MS medium containing 15 g/L sucrose, 1.1 mu M NAA + 2.2 mu M BAP hormone combination, and a pH of 5.7 was highly effective for inducing friable callus from leaf explants of A. malaccensis for the purpose of establishing cell suspension culture.