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      Global gene expression analysis of the shoot apical meristem of maize ( Zea mays L.)

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          Abstract

          All above-ground plant organs are derived from shoot apical meristems (SAMs). Global analyses of gene expression were conducted on maize ( Zea mays L.) SAMs to identify genes preferentially expressed in the SAM. The SAMs were collected from 14-day-old B73 seedlings via laser capture microdissection (LCM). The RNA samples extracted from LCM-collected SAMs and from seedlings were hybridized to microarrays spotted with 37 660 maize cDNAs. Approximately 30% (10 816) of these cDNAs were prepared as part of this study from manually dissected B73 maize apices. Over 5000 expressed sequence tags (ESTs) (about 13% of the total) were differentially expressed ( P<0.0001) between SAMs and seedlings. Of these, 2783 and 2248 ESTs were up- and down-regulated in the SAM, respectively. The expression in the SAM of several of the differentially expressed ESTs was validated via quantitative RT-PCR and/or in situ hybridization. The up-regulated ESTs included many regulatory genes including transcription factors, chromatin remodeling factors and components of the gene-silencing machinery, as well as about 900 genes with unknown functions. Surprisingly, transcripts that hybridized to 62 retrotransposon-related cDNAs were also substantially up-regulated in the SAM. Complementary DNAs derived from the LCM-collected SAMs were sequenced to identify additional genes that are expressed in the SAM. This generated around 550 000 ESTs (454-SAM ESTs) from two genotypes. Consistent with the microarray results, approximately 14% of the 454-SAM ESTs from B73 were retrotransposon-related. Possible roles of genes that are preferentially expressed in the SAM are discussed.

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          Local, efflux-dependent auxin gradients as a common module for plant organ formation.

          Plants, compared to animals, exhibit an amazing adaptability and plasticity in their development. This is largely dependent on the ability of plants to form new organs, such as lateral roots, leaves, and flowers during postembryonic development. Organ primordia develop from founder cell populations into organs by coordinated cell division and differentiation. Here, we show that organ formation in Arabidopsis involves dynamic gradients of the signaling molecule auxin with maxima at the primordia tips. These gradients are mediated by cellular efflux requiring asymmetrically localized PIN proteins, which represent a functionally redundant network for auxin distribution in both aerial and underground organs. PIN1 polar localization undergoes a dynamic rearrangement, which correlates with establishment of auxin gradients and primordium development. Our results suggest that PIN-dependent, local auxin gradients represent a common module for formation of all plant organs, regardless of their mature morphology or developmental origin.
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            The Arabidopsis GNOM ARF-GEF mediates endosomal recycling, auxin transport, and auxin-dependent plant growth.

            Exchange factors for ARF GTPases (ARF-GEFs) regulate vesicle trafficking in a variety of organisms. The Arabidopsis protein GNOM is a brefeldin A (BFA) sensitive ARF-GEF that is required for the proper polar localization of PIN1, a candidate transporter of the plant hormone auxin. Mutations in GNOM lead to developmental defects that resemble those caused by interfering with auxin transport. Both PIN1 localization and auxin transport are also sensitive to BFA. In this paper, we show that GNOM localizes to endosomes and is required for their structural integrity. We engineered a BFA-resistant version of GNOM. In plants harboring this fully functional GNOM variant, PIN1 localization and auxin transport are no longer sensitive to BFA, while trafficking of other proteins is still affected by the drug. Our results demonstrate that GNOM is required for the recycling of auxin transport components and suggest that ARF-GEFs regulate specific endosomal trafficking pathways.
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              DNA sequence quality trimming and vector removal.

              Most sequence comparison methods assume that the data being compared are trustworthy, but this is not the case with raw DNA sequences obtained from automatic sequencing machines. Nevertheless, sequence comparisons need to be done on them in order to remove vector splice sites and contaminants. This step is necessary before other genomic data processing stages can be carried out, such as fragment assembly or EST clustering. A specialized tool is therefore needed to solve this apparent dilemma. We have designed and implemented a program that specifically addresses the problem. This program, called LUCY, has been in use since 1998 at The Institute for Genomic Research (TIGR). During this period, many rounds of experience-driven modifications were made to LUCY to improve its accuracy and its ability to deal with extremely difficult input cases. We believe we have finally obtained a useful program which strikes a delicate balance among the many issues involved in the raw sequence cleaning problem, and we wish to share it with the research community. LUCY is available directly from TIGR (http://www.tigr.org/softlab). Academic users can download LUCY after accepting a free academic use license. Business users may need to pay a license fee to use LUCY for commercial purposes. Questions regarding the quality assessment module of LUCY should be directed to Michael Holmes (mholmes@tigr.org). Questions regarding other aspects of LUCY should be directed to Hui-Hsien Chou (hhchou@iastate.edu).
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                Author and article information

                Journal
                Plant J
                tpj
                The Plant Journal
                Blackwell Publishing Ltd
                0960-7412
                1365-313X
                November 2007
                : 52
                : 3
                : 391-404
                Affiliations
                [1 ]Department of Agronomy, Iowa State University Ames, IA 50011, USA
                [2 ]Bioinformatics and Computational Biology Graduate Program, Iowa State University Ames, IA 50011, USA
                [3 ]Plant Biology Department, University of Georgia Athens, GA 30602, USA
                [4 ]Cold Spring Harbor Laboratory, Cold Spring Harbor NY 11724, USA
                [5 ]Division of Mathematics and Computer Science, Truman State University Kirksville, MO 63501, USA
                [6 ]Division of Science, Truman State University Kirksville, MO 63501, USA
                [7 ]Department of Statistics, Iowa State University Ames, IA 50011, USA
                [8 ]Center for Plant Genomics, Iowa State University Ames, IA 50011, USA
                [9 ]Department of Plant Biology, Cornell University Ithaca, NY 14853, USA
                Author notes
                *(fax +1 515 294 5256; e-mail schnable@ 123456iastate.edu ).
                Article
                10.1111/j.1365-313X.2007.03244.x
                2156186
                17764504
                ef20becb-eae0-4946-8e5b-1210220b3464
                © 2007 The Authors Journal compilation © 2007 Blackwell Publishing Ltd
                History
                : 08 May 2007
                : 31 May 2007
                Categories
                Original Articles

                Plant science & Botany
                shoot apical meristem,global gene expression,development,454 sequencing,retrotransposon expression,laser capture microdissection

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