Anther Culture of Transgenic Pepper (Capsicum annuum L.)

다국어 초록 (Multilingual Abstract)

Pepper (Capsicum annum L.) is an important vegetable crop that can surely benefit from plant biotechnology. Using
transgenic pepper lines resistant to fungal disease, anther culture system was applied to show the possibility to use in molecular
breeding program. We compared some conditions for enhancing anther culture efficiency. We did not find the beneficial effect of
low pretreatment of anthers in both wild type and transgenic lines. By comparing medium compositions, anthers produced more
microspore-derived embryos or embryogenic callus in combination of 0.1 mg/L of 2,4-D and 0.1 mg/L of kinetin. In anther culture
response, wild type had better androgenesis than transgenic lines. PCR results showed that plants regenerated from anthers of
transgenic pepper lines had transgenes, HPT/PepEST. By hygromycin tests, plants developed from anthers of transgenic lines
showed resistance to hygromycin. A total of 48 plants were produced from anther culture of wild type and transgenic lines, among
them, 43 were developed from wild type and 5 plants from transgenic lines. Six plants and one plant spontaneously set pepper
fruits in wild type anther-derived plants and anther derived plants from transgenic lines, respectively. Regenerated plants showed
various phenotypes and some of plants grew normally. These results showed the possibility to use anther culture system for accelerating
breeding programs by the combination with plant biotechnology.

참고문헌 (Reference)

1 "Yang J 1999. Differential interactions of a Colletotrichum gloeosporioides isolate with green and red pepper fruits. Phytoparasitica 27" 97-106,

2 "Tipirdamaz R 2002. The effects of cold treatment and charcoal on the in vitro androgenesis of pepper" 131-139,

3 "Struhl K 1987. Current protocols in Molecular Biology"

4 "Stamp P 2003. Rapid attainment of a doubled haploid line from transgenic maize plants by means of anther culture. In Vitro Cell. Dev. Biol. Plant 39" 165-170,

5 "Skoog F 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15" 473-497,

6 "Pochard E 1981. in vitro culture of pepper high rate plant production from different genotype by +35 treatment. Agronomie 1" 859-864,

7 "Perrier X 2001. Somaclonal variation rate evolution in plant tissue culture Contribution to understanding through a statistical approach. In Vitro Cell. Dev. Biol. Plant 37" 539-542,

8 "Peleman J 2001. QTL mapping of fruit related traits in pepper" 1016-1028,

9 "Nemoto K 2003. Soma-clonal variation of regenerated plants in chili pepper" 233-239,

10 "Lee YI 1975. Studies on the anther culture of Capsicum annuum L. Korean J. Plant Tiss. Cult. 3" 1-7,

11 "Lee YH 2001. Inhibition of fungal appressorium formation by pepper" 80-85,

12 "Kim YS 1998. A microscopic characterization of the infection of green and red pepper fruits by an isolate of Colletotrichum gloeosporioides. J. Phytopathol. 146"

13 "Kim YR 1984. Basic studies on the induction of microspore-originated calluses or embryos in the anther culture of Capsicum annuum L. Korean J. Plant Tiss. cult. 12" 75112-,

14 "Kim M 1999. The influence of temperature pretreatment on the production of microspore embryos in anther culture of Capsicum annuum L. Korean J. Plant Tiss. Cult. 26" 71-76,

15 "Kartha KK 1988. The influence of different temperature treatments on anther culture response of spring wheat" 55-61,

16 "Jang IC 2001. Cytological analysis of microspores during temperature pretreatment in anther culture of Capsicum annuum L. Korean J. Plant Tiss. Cult. 28" 263-271,

17 "In DS 1998. High frequency embryogenesis and plant regeneration in anther cultures of pepper. Kor. J. Plant Tiss. Cult. 25" 319-322,

18 "Guiderdoni E 1996. Transformation of haploid microspore-derived cell suspension protoplasts of rice" 766-770,

19 "Back K 2004. Influence of a selectable marker gene hpt on agronomic performance in transgenic rice. Cereal Res. Comm. 32" 9-16,