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Frontiers of Agriculture in China

ISSN 1673-7334

ISSN 1673-744X(Online)

CN 11-5729/S

Front Agric Chin    2010, Vol. 4 Issue (4) : 438-442     DOI: 10.1007/s11703-010-1028-4
RESEARCH ARTICLE |
Studies on isolated microspore embryoid induction of C. annuum L.
Ting YIN, Song TIAN, Shuangxia LUO, Xueping CHEN, Yanhua WANG, Shuxing SHEN()
College of Horticulture, Agricultural University of Hebei, Baoding 071001, China
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Abstract  

A system to improve isolated microspore embryoid induction rate of pepper (Capsicum annuum L.) was studied in this paper. The results showed that low-temperature pretreatment, growth regulators combinations, activated carbon concentrations, and preculture temperatures were critical factors affecting embryoid formation of isolated pepper microspores in vitro. One day after pretreatment of the buds at 4°C, the anthers that differentiated and developed into embryos were cultured in double-layer medium system for one week at 7°C and then went on culturing at 28°C in darkness. All the seven genotypes of the tested pepper responded to this protocol. The embryoid induction rate of the best genotype increased from 81.11% to 147.22%. This protocol can be used as a potential tool for producing doubled haploid plants for pepper breeding.

Keywords Capsicum annuum L.      isolated-microspore culture      embryoids      embryoid induction rate      double-layer medium system     
Corresponding Authors: SHEN Shuxing,Email:shensx@hebau.edu.cn   
Issue Date: 05 December 2010
URL:  
http://academic.hep.com.cn/fag/EN/10.1007/s11703-010-1028-4     OR     http://academic.hep.com.cn/fag/EN/Y2010/V4/I4/438
serial number1234
ZT/mg·L-10.50.51.01.0
IAA/mg·L-10.51.00.51.0
Tab.1  Combinations of growth regulators
Fig.1  Time-course of events during isolated microspore culture of pepper
Note: (a): late-unicellular microspores; (b): cell clusters after 3 weeks of microspore culture; (c) and (d): categories of embryos obtained from microspore culture after 7 weeks; (e) and (f): differentiated embryoid of microspore culture; (g) and (h): regenerated plantlets.
serial numbergenotypesembryos inducedembryoid induction rate /%
108-2002319.44cd
208-2042210.56d
308-2349942.78b
408-2648235.56b
508-48819281.11a
608-490509.44d
708-4914423.89c
Tab.2  Embryos induced from different genotypes under the same cultural conditions
serial numberCK1234
different periods of 4°C cold pretreatment of the flower buds/d01234
induction rate of embryos induced/%79.63b106.48a87.96b64.81c45.37d
Tab.3  Effect of low-temperature pretreatment of the flower buds for the embryoids induction rate
serial numbertemperature during the first week/°Cembryoids induction rate /%combination ofgrowth regulators/mg·L-1embryoids induction rate /%
CK2819.44e082.41b
1470.37b0.5ZT+ 0.5IAA57.41c
27100.93a0.5ZT+ 1.0IAA147.22a
33252.85c1.0ZT+ 0.5IAA42.59c
43535.18 d1.0ZT+ 1.0IAA25.93d
Tab.4  Effect of preculture temperature during the first week and the combinations and concentrations of growth regulators on embryoids induction rate
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