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Published on February 19, 2004; 10.1104/pp.103.035998

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Received November 11, 2003
Returned for revision December 4, 2003
Accepted December 14, 2003

A Novel Endo-{beta}-Mannanase Gene in Tomato LeMAN5 Is Associated with Anther and Pollen Development

Sergei A. Filichkin , Jeffrey M. Leonard , Alvaro Monteros , Po-Pu Liu , and Hiroyuki Nonogaki *

Departments of Horticulture (S.A.F., A.M., P.-P.L., H.N.) and Environmental and Molecular Toxicology (J.M.L.), Oregon State University, Corvallis, Oregon 97331-7304

* Corresponding author; email: hiro.nonogaki{at}oregonstate.edu.

Endo-{beta}-mannanase (EC 3.2.1.78) is involved in cell wall disassembly and the weakening of plant tissues by degrading mannan polymers in the cell walls. Endo-{beta}-mannanase genes are expressed in tomato (Lycopersicon esculentum) seeds (LeMAN1 and LeMAN2) and fruits (LeMAN3 and LeMAN4). A novel endo-{beta}-mannanase gene (termed LeMAN5) was found in the tomato genome by genome-walking PCR and bacterial artificial chromosome library screening. The 5'-upstream region of this endo-{beta}-mannanase gene contained four copies of the pollen-specific cis-acting elements POLLEN1LELAT52 (AGAAA). A GUS-reporter gene driven with the putative LeMAN5 promoter (-543 to +38) was activated in anthers and pollen of transgenic Arabidopsis, with the highest {beta}-glucuronidase activity detected in pollen. {beta}-Glucuronidase expression was detected in mature pollen retained in sporangia, discharged pollen, and elongating pollen tubes in transgenic Arabidopsis. Consistently, expression of LeMAN5 mRNA and endo-{beta}-mannnanase activity was detected in tomato anthers and pollen. In anthers, the highest mRNA expression and endo-{beta}-mannanase activity were detected during late stages of anther development, when pollen maturation occurred. Endo-{beta}-mannanase activity was present in discharged pollen, which was easily eluted in a buffer, indicating that the enzyme proteins are probably secreted from, and deposited on, the surface of pollen. These data suggest that the LeMAN5 endo-{beta}-mannanase is associated with anther and pollen development.




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