Expression and regulation of two novel anther-specific genes in Lilium longiflorum.
Two stage-specific genes have been isolated from a subtractive cDNA library constructed from developing anthers of lily (Lilium longiflorum). The proteins encoded by the two genes have a strong hydrophobic region at the N-terminus, indicating the presence of a signal peptide. The deduced LLA-67 is a new type of small cysteine-rich protein whose sequence exhibits four consecutive CX(3)CX(6-10) repeats that could form signal-receiving finger motifs, while the deduced LLA-115 protein shows significant similarities to a rice unknown protein, and putative cell wall proteins of Medicago truncatula and Arabidopsis. The transcripts of LLA-67 and LLA-115 were anther specific and differentially detected at the phase of microspore development. In situ hybridization with antisense riboprobes of the two genes in the anther showed strong signals localized to the tapetal layer of the anther wall. The LLA-67 mRNA was also detected in the microspore at the phase of microspore development but the LLA-115 mRNA was not. The LLA-115 gene can be exogenously induced by gibberellin (GA), whereas the LLA-67 gene cannot be induced. Studies with the GA biosynthesis inhibitor uniconazole and an inhibitor of ethylene activity, 2,5-norbornadien (NBD), revealed that the two genes were negatively regulated by ethylene and a cross-talk between GA and ethylene was involved in the regulation of the two genes occurring in young anthers. The treatment of NBD caused the tapetum to become densely cytoplasmic and highly polarized, whereas uniconazole arrested tapetal development to a status close to that of control. DNA blots of lily genomic DNA indicated that the two genes were encoded by a small gene family. The different actions of hormones on gene expression and the possible function of the gene products in young anthers are discussed.
Tzeng
Jhih-Deng
JD
Graduate Institute of Biotechnology, National Chung Hsing University, Taichung, Taiwan.
Hsu
Ssu-Wei
SW
Chung
Mei-Chu
MC
Yeh
Fung-Ling
FL
Yang
Chin-Ying
CY
Liu
Ming-Che
MC
Hsu
Yi-Feng
YF
Wang
Co-Shine
CS
D000595:Amino Acid Sequence; D018076:DNA, Complementary / Q000235:genetics; D018744:DNA, Plant / Q000235:genetics; D005030:Ethylenes / Q000494:pharmacology; D020869:Gene Expression Profiling; D018506:Gene Expression Regulation, Plant / Q000187:drug effects; D017343:Genes, Plant; D005875:Gibberellins / Q000494:pharmacology; D027762:Lilium / Q000166:cytology / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development; D008969:Molecular Sequence Data; D009928:Organ Specificity / Q000187:drug effects / Q000235:genetics; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D011058:Pollen / Q000166:cytology / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D013997:Time Factors
D018076:DNA, Complementary; D018744:DNA, Plant; D005030:Ethylenes; D005875:Gibberellins; D010940:Plant Proteins; D012333:RNA, Messenger; C036216:ethylene; C007842:gibberellic acid