Transgenic Tobacco Plants Expressing Pea Chloroplast Nmdh cDNA in Sense and Antisense Orientation (Effects on NADP-Malate Dehydrogenase Level, Stability of Transformants, and Plant Growth)

Abstract

A full-length cDNA encoding light-activated chloroplast NADP-malate dehydrogenase (NADP-MDH) (EC 1.1.1.82) from pea (Pisum sativum L.) was introduced in the sense and antisense orientation into tobacco (Nicotiana tabacum L.). Transgenic plants with decreased or increased expression levels were obtained. Because of substantial age-dependent differences in individual leaves of a single plant, standardization of NADP-MDH levels was required first. Then, extent and stability of over- or under-expression of Nmdh, the gene encoding NADP-MDH, was characterized in the various transformants. Frequently, cosuppression effects were observed, indicating sufficient homology between the endogenous tobacco and the heterologous pea gene. Analysis of the T1 and T2 progeny of a series of independent transgenic lines revealed that NADP-MDH capacity ranged between 10% and [greater than or equal to]10-fold compared with the wild type. Under ambient conditions whole-plant development, growth period, and fertility were unaffected by NADP-MDH reduction to 20% of the wild-type level; below this threshold plant growth was retarded. A positive growth effect was registered in young plants with stably enhanced NADP-MDH levels within a defined developmental window.