Protein Analysis of Successive Continuous Generation Cuttings of Buxus Sinica Var.Parvifolia

Advance in Forestry Research

November 2013, Volume 1, Issue 4, PP.45-50 Protein Analysis of Successive Continuous Generation Cuttings of Buxus Sinica

Var.Parvifolia

Kongshu Ji #, Mingjuan Wang, Yan Huang, Xiaofeng Wang, Ting Pan

1. Ministry of Education, Nanjing Forestry University, Nanjing, 210037, P.R. China

#Email: ksji@

Abstract

Buxus sinica var. parvifolia is a rare endangered and endemic tree species in China, as well as a kind of precious material for bonsai. The experimental materials were leaves of B. sinica var. Parvifolia collected from seedling planted in 2003 and clonal stecklings of three successive generations raised in 1994, 1998 and 2000 respectively. The proteins of the materials were extracted with the 2-D Clean-Up Kit and analyzed with two dimensional gel electrophoresis. The gels were then analyzed by Image Master TM 2D Platinum 6.0 software. Four hundred and fifty-five reproducible protein spots were detected, among which forty-nine protein spots displayed differential expression. There were twenty-five, nineteen and twelve different protein spots between seedling in 2003 and steckling of generation 1994, steckling of generation1994 and 1998, and steckling of generation 1998 and 2000 respectively. Among all of them, eight proteins were up-regulated and six proteins were down-regulated. These proteins may be related to rejuvenation of Buxus sinica var. parvifolia.

Keywords: B. Sinica Var. Parvifolia; Rejuvenation; Two Dimensional Gel Electrophoresis; Differential Protein

1I NTRODUCTION

Buxus. sinica var. parvifolia is a rare and endangered tree species distributing in some subtropical and alpine areas of China and plays an important role in the maintenance of the landscape and ecosystem in its cliff habitats at elevations between 1200 m and 3000 m above sea level. It is an excellent material to manufacture bonsai. The successive generation cutting method is used to instruct and domesticate Buxus. sinica var. parvifolia until it completely retains its original morphological characteristics and gradually adapts to current cultivation environment (Huang, 2008).

“Rejuvenation” is usually used to describe the phenomenon that tree organs or cell formations recover their juvenile characteristics, such as strong rooting ability, flourish leaves, stronger cold resistance and stronger drought resistance. Asexual rejuvenation of the tree occurs when its somatic cells receive specific exogenous stimulation or gene activation. mRNA or proteins might act as the provider of rejuvenation signals(Chen, 1996). The recent researches showed more ways about how to rejuvenate but less on its mechanism. The mechanism of rejuvenation was analyzed through studying the proteins of successive generation cuttings of Buxus sinica var. parvifolia with two-dimensional gel electrophoresis (2-DE), which also provides help for the proteome research of B. sinica var. parvifolia.

2M ATERIALS AND M ETHODS

2.1 Materials

The leaves of B. sinica var. parvifoliawere were collected from seedling and successive generations of stecklings in Haoyu Landscaping Company, Rugao, Jiangsu Province, and China. The seedling were planted in 2003 and referred to as seedling in 2003 or seedling. The successive generations of stecklings of Buxus sinica var. parvifolia were obtained through the following method: in 1990, the first generation twigs were taken as cuttings from one original