Plant and Cell Physiology Advance Access originally published online on September 20, 2006
Plant and Cell Physiology 2006 47(11):1457-1472; doi:10.1093/pcp/pcl013
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Rice Immature Pollen 1 (RIP1) is a Regulator of Late Pollen Development
1National Research Laboratory of Plant Functional Genomics, Division of Molecular and Life Sciences, Republic of Korea
2Functional Genomic Center, Pohang University of Science and Technology, Pohang 790-784, Republic of Korea
3Yeongnam Agricultural Research Institute, National Institute of Crop Science, Rural Development Administration, Milyang, 627-130, Republic of Korea
* Corresponding author: E-mail, genean{at}postech.ac.kr; Fax, +82-54-279-0659.
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Abstract |
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We isolated a pollen-preferential gene, RICE IMMATURE POLLEN 1 (RIP1), from a T-DNA insertional population of japonica rice that was trapped by a promoterless ß-glucuronidase (GUS) gene. Semi-quantitative reverse transcription–PCR (RT–PCR) analyses confirmed that the RIP1 transcript was abundant at the late stages of pollen development. Transgenic plants carrying a T-DNA insertion in the RIP1 gene displayed the phenotype of segregation distortion of the mutated rip1 gene. Moreover, rip1/rip1 homozygous progeny were not present. Reciprocal crosses between Rip1/rip1 heterozygous plants and the wild type showed that the rip1 allele could not be transmitted through the male. Microscopic analysis demonstrated that development in the rip1 pollen was delayed, starting at the early vacuolated stage. Close examination of that pollen by transmission electron microscopy also showed delayed formation of starch granules and the intine layer. In addition, development of the mitochondria, Golgi apparatus, lipid bodies, plastids and endoplasmic reticulum was deferred in the mutant pollen. Under in vitro conditions, germination of this mutant pollen did not occur, whereas the rate for wild-type pollen was >90%. These results indicate that RIP1 is necessary for pollen maturation and germination. This gene encodes a protein that shares significant homology with a group of proteins containing five WD40 repeat sequences. The green fluorescent protein (GFP)–RIP1 fusion protein is localized to the nucleus. Therefore, RIP1 is probably a nuclear protein that may form a functional complex with other proteins and carry out essential cellular and developmental roles during the late stage of pollen formation.
Keywords: Male gametogenesis - Nuclear localization - Pollen maturation - Rice - T-DNA gene trap
Abbreviations: DAPI, 4',6-diamidino-2-phenylindole; GFP, green fluorescent protein; GUS, ß-glucuronidase; RFP, red fluorescent protein; RT–PCR, reverse transcription–PCR; TAIL-PCR, thermal asymmetric interlaced PCR.
4 Present address: Department of Plant Pathology, University of California, Davis, CA 95616, USA.
(Received May 29, 2006; Accepted September 14, 2006)
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