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DEVELOPMENT AND HORMONE ACTION

A Novel Endo--Mannanase Gene in Tomato LeMAN5 Is Associated with Anther and Pollen Development¹

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

Endo--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--mannanase genes are expressed in tomato (Lycopersicon esculentum) seeds (LeMAN1 and LeMAN2) and fruits (LeMAN3 and LeMAN4). A novel endo--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--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 -glucuronidase activity detected in pollen. -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--mannnanase activity was detected in tomato anthers and pollen. In anthers, the highest mRNA expression and endo--mannanase activity were detected during late stages of anther development, when pollen maturation occurred. Endo--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--mannanase is associated with anther and pollen development.

¹ This work was supported by the National Science Foundation (grant no. IBN–0237562).

² Present address: Department of Crop and Soil Science, Crop Science Building, Oregon State University, Corvallis, OR 97331–3002.

³ Present address: Departamento Nacional de Recursos Fitogeneticos y Biotecnologia, Instituto Nacional Autonomo de Investigaciones Agropecuarias, Estacion Experimental Santa Catalina, P.O. Box 17–01–340, Quito, Ecuador.

^* Corresponding author; e-mail hiro.nonogaki{at}oregonstate.edu; fax 541–737–3479.

Received November 11, 2003; returned for revision December 4, 2003; accepted December 14, 2003.

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