Figure 4.

Characterization of the lhca6 RNAi Lines.

(A) RT-PCR analysis of Lhca6 and Lhca2 mRNA. Total RNA (5 μg) was reverse transcribed, and the resulting cDNA was used in 30 cycles of PCR with specific primers for Lhca6, Os Lhca6, and Lhca2. ACT8 was used as an internal control. “lhca6c” indicates lhca6 RNAi mutant complemented by the introduction of Os Lhca6 gene.

(B) Monitoring of NDH activity by chlorophyll fluorescence as in Figure 1C.

(C) Immunodetection of chloroplast proteins from immature and mature leaves of the wild type and lhca6. The thylakoid membrane proteins were separated by SDS-PAGE and immunodetected with specified antibodies. Thylakoid proteins were loaded on an equal chlorophyll basis. These experiments were repeated three times independently, and similar results were obtained. Results from a representative experiment are shown.

(D) Analysis of thylakoid proteins. Immunoblot results of three independent isolations of thylakoid membranes were analyzed with Imagemaster software (Amersham Pharmacia Biotech). The protein levels in the wild-type mature leaves and lhca6 immature and mature leaves are shown relative to those in the wild-type immature leaves (100%). Means ± sd (n = 3).