《植物生理学报》 2017, 53(3): 422-428

日本五针松针叶胚性愈伤诱导及愈伤组织细胞学特性观察

王雪丽, 李萍萍, 吴坤阳, 石琨, 郑彩霞^*

北京林业大学生物科学与技术学院, 北京100083

通信作者：郑彩霞；E-mail: zhengcx@bjfu.edu.cn

摘　要：

为建立日本五针松再生体系, 本文以其幼嫩针叶为材料, 在DCR基本培养基上分别诱导愈伤组织和胚性愈伤组织, 探索其最佳植物生长调节物质配比, 并在诱导过程中对愈伤组织进行细胞学特性观察。结果表明, 使用添加5 mg·L^-1 2,4-D和1 mg·L^-1 6-BA的培养基, 愈伤组织的诱导率高达96.7%。胚性愈伤诱导培养基中添加0.6 mg·L^-1 TDZ、1.0 mg·L^-1复硝酚钠、1.0 mg·L^-1 6-BA和0.2 mg·L^-1 IBA, 可诱导细胞分化, 获得胚性愈伤, 诱导率可达53.3%, 而且其褐化率低, 生长较快。非胚性愈伤的细胞呈圆形和细长形两种, 不形成细胞团; 胚性愈伤的细胞结合紧密, 形成了胚性细胞团, 其主要由胚性细胞和胚柄细胞两种类型的细胞组成。研究结果为日本五针松再生体系建立奠定了重要的基础。

关键词：日本五针松; 非胚性愈伤; 胚性愈伤; 形态学与细胞学特性

收稿：2016-12-15   修定：2017-01-16

资助：国家自然科学基金(31170563)。

Inducement of embryonic callus from needles of Pinus parviflora Sieb. et Zucc. and observation of cytological characteristics of callus

WANG Xue-Li, LI Ping-Ping, WU Kun-Yang, SHI Kun, ZHENG Cai-Xia^*

College of Biological Science and Technology, Beijing Forest University, Beijing 100083, China

Corresponding author: ZHENG Cai-Xia; E-mail: zhengcx@bjfu.edu.cn

Abstract:

In order to establish the regeneration system Pinus parviflora Sieb. et Zucc., the young needles were used to induce callus and embryonic callus in DCR medium and the best plant stimulus ratio was explored. In the process of induction, the tissues were observed by optical microscope and scanning electron microscope. The results indicated that the induction rate of callus was high as 96.7% with the addition of 5 mg·L^-1 2,4-D and 1 mg·L^-1 6-BA. Embryonic callus induction medium, including 0.6 mg·L^-1 TDZ, 1.0 mg·L^-1 compound sodium nitrophenolate, 1.0 mg·L^-1 6-BA and 0.2 mg·L^-1 IBA, could induce cell differentiation and obtain embryonic callus. The induction rate reached 53.3%. At the same time, the browning rate of embryonic callus was low and the tissues grew faster. The cells of the non-embryonic callus were round and thin, and no cell clusters were formed. The cells of embryonic callus bound with each other tightly and formed the embryonic cell clusters which were mainly composed of embryogenic cells and suspensor-like cells. These results laid an important foundation for the establishment of regeneration system for P. parviflora.

Key words: Pinus parviflora Sieb. et Zucc.; non-embryonic callus; embryonic callus; morphological and cytological characteristics