THE COMPETENCE OF CELLS FOR CELL-DIVISION AND REGENERATION IN TOBACCO EXPLANTS DEPENDS ON CELLULAR LOCATION, CELL-CYCLE PHASE AND PLOIDY LEVEL

被引:24
作者
GILISSEN, LJW
VANSTAVEREN, MJ
HAKKERT, JC
SMULDERS, MJM
VERHOEVEN, HA
CREEMERSMOLENAAR, J
机构
[1] Department of Cell Biology, DLO-Centre for Plant Breeding and Reproduction Research (CPRO-DLO), NL-6700 AA Wageningen
关键词
CELL CYCLE; FLOW CYTOMETRY; CONFOCAL LASER SCANNING MICROSCOPY; REGENERATION; THIN CELL-LAYER (TCL) EXPLANT; NICOTIANA TABACUM;
D O I
10.1016/0168-9452(94)03976-3
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
This work concerns application of flow cytometry (FCM) and confocal laser scanning microscopy (CLSM) to investigate the competence of cells for cell division and regeneration. FCM analysis of freshly-cut thin cell layer (TCL) explants of Nicotiana tabacum, excised from upper internodes of vegetative plants, revealed that one-quarter of the cells had the 2C nuclear DNA content, whereas of the other cells most nuclei had the 4C and some had the 8C DNA content. Cytometric examination using CLSM showed that the 2C nuclei were mainly located in the epidermis and subepidermis, and the 4C nuclei predominantly in the cortical tissue. During culture of the explants, part of the cortical cells went through mitosis from the first day onwards, and formed callus from which predominantly diploid and some tetraploid roots regenerated at low frequency. Most cortical cells were thus in the G(2) phase of the diploid cell cycle. FCM analysis showed that another fraction of the 4C cortical cells was induced to endoreduplicate to 8C cells. These cells thus had previously switched to the G(1) phase of the tetraploid cell cycle. CLSM analysis revealed that subepidermal and epidermal cells, respectively, underwent cell division from the second and third day onwards. Shoot primordia developed from cells of both cell layers together. Most shoot regenerants were normal diploids, some were mixoploids or tetraploids. The combination of FCM and CLSM allowed identification of the cell cycle phase, the ploidy level, the position of the cell, and the cellular development. The results give insight into the involvement of these parameters in the competence for cell division and regeneration at the level of the individual explant cells, and are therefore relevant for cellular and molecular approaches to plant transformation.
引用
收藏
页码:81 / 91
页数:11
相关论文
共 34 条
[1]  
ALTAMURA MM, 1984, ANN BOT ROMA, V42, P45
[2]  
BROWN SC, IPATIMUP SERIES, V1
[3]  
COUSSON A, 1992, PHYSIOL PLANTARUM, V85, P102, DOI 10.1111/j.1399-3054.1992.tb05270.x
[4]   HISTOLOGY OF THE MORPHOGENIC RESPONSE IN THIN CELL LAYER EXPLANTS FROM VEGETATIVE TOBACCO PLANTS [J].
CREEMERSMOLENAAR, J ;
HAKKERT, JC ;
VANSTAVEREN, MJ ;
GILISSEN, LJW .
ANNALS OF BOTANY, 1994, 73 (05) :547-555
[5]   CYTOGENETICS OF PLANT-CELL AND TISSUE-CULTURES AND THEIR REGENERATES [J].
DAMATO, F .
CRC CRITICAL REVIEWS IN PLANT SCIENCES, 1985, 3 (01) :73-112
[6]   FATE OF INTRODUCED GENETIC-MARKERS IN TRANSFORMED ROOT CLONES AND REGENERATED PLANTS OF MONOHAPLOID AND DIPLOID POTATO GENOTYPES [J].
DEVRIESUIJTEWAAL, E ;
GILISSEN, LJW ;
FLIPSE, E ;
RAMULU, KS ;
STIEKEMA, WJ ;
DEGROOT, B .
THEORETICAL AND APPLIED GENETICS, 1989, 78 (02) :185-193
[7]  
EBERHARD S, 1989, PLANT CELL, V1, P747, DOI 10.1105/tpc.1.8.747
[8]   RAPID FLOW CYTOMETRIC ANALYSIS OF THE CELL-CYCLE IN INTACT PLANT-TISSUES [J].
GALBRAITH, DW ;
HARKINS, KR ;
MADDOX, JM ;
AYRES, NM ;
SHARMA, DP ;
FIROOZABADY, E .
SCIENCE, 1983, 220 (4601) :1049-1051
[9]   SYSTEMIC ENDOPOLYPLOIDY IN ARABIDOPSIS-THALIANA [J].
GALBRAITH, DW ;
HARKINS, KR ;
KNAPP, S .
PLANT PHYSIOLOGY, 1991, 96 (03) :985-989
[10]   DEVELOPMENT OF POLYSOMATY IN SEEDLINGS AND PLANTS OF CUCUMIS-SATIVUS L [J].
GILISSEN, LJW ;
VANSTAVEREN, MJ ;
CREEMERSMOLENAAR, J ;
VERHOEVEN, HA .
PLANT SCIENCE, 1993, 91 (02) :171-179