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Title: Structure and expression of ribulose-1, 5-biphosphate carboxylase/oxygenase small subunit genes in sugarcane 
Author: Tang, Wendong
Date: 1994
Abstract: Sugarcane (Saccharum spp. hybrid cultivar H32-8560), a C4 polyploid, is estimated to contain about 16 copies of the ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit (rbcs) gene. Seven rbcs genes were isolated from a sugarcane genomic library, and were partially or completely sequenced. Unlike rbcs gene members of other plant species that are heterogeneous in their 3' UTRs, these genes are highly conserved in their coding sequences as well as their 3' and 5' untranslated regions (UTRs). The nested Rapid Amplification of cDNA 3' Ends (nested-RACE) technique was used to clone the 3' UTR of rbcs mRNA from sugarcane leaves of different developmental stages. There is no significant difference among the RACE-products from different organs in terms of their overall electrophoretic patterns. The sequences of the 3 '-RACE products have more than 95% homology with the 3' UTR of the cloned rbcs genes. However, their lengths (from translation stop codon to polyadenylation site) are quite different and can be classified into five major size groups, i.e. Group I, 501bP; II, 351bP; III, 254bP; VI, 184bP; and V, 103bp. Northern analysis of total RNA from mature leaf, 51 mapping, and sequence determination of the rbcs cDNA clones isolated from a mature leaf cDNA library confirm the existence of different lengths of rbcs mRNA. The AAUAAA-like motifs for Groups I and II can be identified; however, we fail to detect other sequence motifs that have been suggested to be important in directing correct cleavage and polyadenylation. Differential amplification of sugarcane genomic DNA suggests that the genes encoding Groups I and II mRNAs are similar in length. Groups I and II mRNAs continue to be expressed during development of the sugarcane leaf and the ratio of these two classes of mRNAs in various sugarcane samples is similar. Sugarcane leaf bombardment experiments showed that both the scrbcs-1 and scrbcs-3 promoters are able to direct GUS gene expression in mesophyll cells and bundle sheath cells during C3-C4 transition, although at a ratio of 1 to 3 in favor of the bundle sheath cells. These finding supports the notion that the rbcs genes in C4 plants have a default C3-type expression pattern (Schaffner and Sheen, 1991).
Description: Thesis (Ph. D.)--University of Hawaii at Manoa, 1994. Includes bibliographical references (leaves 126-140). Microfiche. 140 leaves, bound ill., photographs (some col.) 29 cm
URI: http://hdl.handle.net/10125/9476
Rights: All UHM dissertations and theses are protected by copyright. They may be viewed from this source for any purpose, but reproduction or distribution in any format is prohibited without written permission from the copyright owner.
Keywords: Sugarcane -- Genetics

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