湿地松细胞系内源激素及生理生化特性与成胚能力相关性研究

Correlation study between endogenous hormones and physiological and biochemical properties and somatic embryogenic capacity of Pinus elliottii cell lines

关键词:  湿地松, 细胞系, 生理生化, 内源激素, 成胚能力

Key words:  Pinus elliottii, cell line, physiological and biochemical, endogenous hormones, embryogenic capacity

摘要:

为探究影响细胞系胚性强弱的特征指标值,早期判别胚性较强的胚性细胞系。该研究以湿地松未成熟种子诱导的7个细胞系为材料,观测其表型特征和细胞结构,测定内源激素含量、可溶性蛋白含量、可溶性糖含量、淀粉含量、抗氧化酶活性等指标,并选择5个细胞系进行成熟培养,采用方差分析、Duncan多重比较及Pearson相关性分析,研究了湿地松不同细胞系内源激素含量及生理生化特性与成胚能力的相关性。结果表明:(1)不同细胞系在细胞学观察上存在差异,其中V30细胞系胚性胚柄细胞团结构明显、数量较多、胚性较强。(2)不同细胞系的内源激素含量存在显著性差异,脱落酸(ABA)、玉米素核苷(ZR)与细胞系成胚能力存在显著正相关,吲哚乙酸(IAA)含量与细胞系成胚能力存在显著负相关,赤霉素(GA₃)差异不显著且与细胞系成胚能力无明显相关性。(3)不同细胞系的可溶性蛋白含量、可溶性糖含量、淀粉含量、过氧化物酶(POD)、超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性均存在显著性差异,但与细胞系成胚能力无明显相关性。(4)以细胞系胚性和成熟胚数量为标准,V30细胞系具有高体胚发生潜力。综上认为,湿地松细胞系的成胚能力与内源激素含量显著相关,这不仅为进一步通过调节激素含量来改善细胞系胚性和优化体胚成熟条件提供了参考与借鉴,而且还为优化湿地松体胚发生和提高体胚诱导率等方面的研究提供了技术参考。

Abstract:

To study the correlation between endogenous hormone content and physiological and biochemical characteristics and somatic embryogenic capacity for different cell lines of Pinus elliottii, seven cell lines induced by immature seeds of P. elliottii were used as materials. The phenotype and cellular structure were observed, and the contents of endogenous hormones, soluble protein, soluble sugar, starch and antioxidant enzyme activities were determined, followed by the maturation culture of five selected cell lines. In order to explore the characteristic index values that affect the embryogenic capacity of cell lines, so as to identify the embryogenic capacity of cell lines with strong embryogenic capacity in the early stage, and provide technical reference for optimizing the somatic embryogenesis and improving the somatic embryo induction rate of P. elliottii. Data analysis involved analysis of variance(ANOVA), Duncan multiple range test, and pearson correlation analysis. The results were as follows:(1)Different cell lines differed in cytological observation, among which the V30 cell line had obvious and more embryonal-suspensor mass(ESM), indicating the V30 line was a strong embryonic line.(2)The endogenous hormone content was significantly and different among cell lines, with a significant positive correlation among abscisic acid(ABA), trans-zeatin riboside(ZR)and cell lines, a significant negative correlation among indoleacetic acid(IAA)content and cell lines, no significant difference among gibberellic acid(GA₃)and no significant correlation among GA₃ and cell lines.(3)The soluble protein content, soluble sugar content, starch content, peroxidase(POD), superoxide dismutase(SOD), and catalase(CAT)activity were significantly different among cell lines, but showed no obvious correlation with the embryogenic ability of cell lines.(4)Based on the embryogenesis of the cell line and the number of mature embryos, the V30 cell line had a high somatic embryogenesis potential. The somatic embryogenic capacity of the P. elliottii cell line is significantly correlated with the endogenous hormone content, and it can provide reference for improving the embryogenic capacity of cell lines and optimizing the maturation conditions of somatic embryos.