Production and Characterization of Nitrate Reductase Deficient Mutants in Petunia parviflora

• 영문명: Production and Characterization of Nitrate Reductase Deficient Mutants in Petunia parviflora
• 발행기관: 한국자원식물학회
• 저자명: Lee, Cheol-Hee
• 간행물 정보: 『한국자원식물학회지』제19권 제6호, 706~715쪽, 전체 10쪽
• 주제분류: 농수해양 > 기타농수해양
• 파일형태: PDF
• 발행일자: 2006.12.30

영문 초록

Nitrate reductase deficient (NR) mutant lines were selected indirectly by their resistance to 100mM chlorate in cell cultures of P. parviflora. A total of 585 chlorate resistant lines were confirmed by a second passage on a high concentration of chlorate. Frequency of spontaneous mutation was $9.7{\times}10^{-7}$ 수식 이미지 in 3 month old suspension-cultured cells, and in non-selective media containing amino acids as sole nitrogen source. The frequency of mutation could be increased up to 11-fold by culture for 12 months. Out of 40 randomly selected calli, 22 were fully deficient in NR. The rest of the clones contained a decreased level of NR activity. Further characterization was carried out in 13 mutant lines which were fully deficient in NR and in 5 mutant lines containing residual (0-7.0%) NR activity, as compared to wild-type cells cultured on the same medium. The $NR^-$ 수식 이미지 mutants were tentatively classified as defective in the NR apoenzyme (nia-type; 11 mutant lines including the 5 with residual NR activity) or in the molybdenum cofactor (cnx-type; 7 mutant lines) by the XDH activity. The cnx-type could be further classified into two groups. In one group (5 mutant lines) of these, the NR activity could be partially restored by nonphysiologically high (1.0mM) molybdate in the culture medium. Both types of $NR^-$ 수식 이미지 mutants were unable to grow on minimal medium containing nitrate as sole nitrogen source, but grew well on amino acids. They also proved to be extremely sensitive to the standard medium ($MSP_1$ 수식 이미지) containing nitrate and ammonium. Shoot regeneration was obtained only in the $NR^-$ 수식 이미지 mutants, which contained residual NR activity, but they so far have failed to grow into plants.

키워드

Nitrate reductase deficient ( NR− ) mutant Petunia parviflora NR apoenzyme Molybdenum cofactor