EST analysis in Calamus manan MIQ

For the first time, eight floral cDNA libraries (four male viz. ECM01, ECM02, ECM03, ECM04 and four female viz. ECF01, ECF02, ECF03, ECF04) were constructed from different developmnental stages of male and female inflorescences, leaf (ECL01) and fruit (ECFR01) of Calamus manan. The floral libraries were generated to identify floral ESTs that may be developed into DNA markers for sex determination in C manan. The insert sizes of cDNA clones from ECF02 (18,000 clones), ECM02 (6,359 clones), ECM01 (5,000 clones), ECF01 (1,179 clones), ECF03 (5,000 clones) ECF04 (9,108 clones), ECM03 (5,980 clone), ECM04 (5,318 clones), ECL01 (3.840 clones) and ECFR01 (768 clones) ranged from 0.5 to 2.5 kb with an average insert size of 1.0 kb. A total of 1529 floral ESTs generated randomly from the above floral libraries were analyzed and compared against 371 fruit ESTs and 338 leaf ESTs. The floral ESTs were grouped into 4 categories in term of protein matching: (1) significant with known function (49.89%), (2) significant with unknown function & hypotheticalproteins (78%). (3) not significant (34.1%) and (4) no match (8.3%). Further analysis on floral ESTs from group 1 yielded 8 categories of functional genes metabolism (30.4%), protein synthesis (28.0%), organization and cellular function (23.8%), defense and stress regulation (12.6%), flowering (0.4%), development (1.4%) and miscellaneous (3.4%). Meanwhile a major portion of the leaf ESTs (29.7%) were involved in photosynthesis and most of the fruit ESTs had unknown gene function (27%). In addition, 11.3% of the ESTs generated from the fruit, floral and leaf cDNA showed no significant homology to any entry in the Genbank. A total of 3.3% of group 1 and 2 floral ESTs were floral genes. The floral genes identified were the MADS box gene 8, protein stamen specific fil1, CONSTANS, FRIGIDA stigma/stylar cysteine-rich adhesion precursor, flower-specific gamma thionine precursor, anther- specific proline-rich protein and Early Flowering 5.