Different gene expression patterns of sucrose-starch metabolism during pollen maturation in cytoplasmic male-sterile and male-fertile lines of rice

Starch biosynthesis, a very critical event during pollen development, is not well understood in terms of gene regulation and intracellular controls. Using the rice HL-type cytoplasmic male sterile (HL-CMS) line with starch deficiency in pollen and two related male-fertile lines (Oryza sativa L.), we have studied two specific developmental stages: 'early', that is, before or during pollen mitosis I, without starch accumulation, and 'late', the actively starch-filling postpollen mitosis I phase. Real-time reverse transcription-PCR analysis showed temporal changes in the expression patterns of six metabolic genes involved in the sucrose-starch pathway, i.e. sucrose synthase (EC 2.4.1.13), Rsus3; acid invertase (EC 3.2.1.26), OSINV2; phosphoglucomutase (EC 5.4.2.2), OsPGM; uridine diphosphate-glucose pyrophosphorylase (EC 2.7.7.9), OsUGP; a subunit of adenosine diphosphate-glucose pyrophosphorylase (AGPase) (EC 2.2.7.2), OsAGPL3; soluble starch synthase (EC 2.4.1.21), OsSSI and three transporter genes [sucrose transporter, OsSUT3; hexose transporter, OSMST7 and plasma membrane H+-adenosine triphosphatase (ATPase) (EC 3.6.1.35), OSA2]. Regarding metabolic enzyme activities, lower activity of sucrose synthase was detected at 'late' compared with 'early' stage in the three rice lines; invertase and other downstream enzymes had higher activities at 'late' stage in male-fertile but not male-sterile lines. Meanwhile, significantly reduced sugars and ATP levels were detected at 'early' and 'late' stages in male-sterile compared with male-fertile tissues. Furthermore, transgenic data showed that AGPase gene can stimulate starch accumulation in pollen of male-sterile host lines, showing the importance of AGPase in the metabolic pathway. Taken together, these data suggest that the combined effects of the reduced energy supply and sugar levels in starch biosynthesis may lead to the observed temporal changes in gene expression, and eventually to pollen sterility.