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US 20200407740A1 IN ( 19 ) United States ( 12 ) Patent Application Publication ( 10 ) Pub . No .: US 2020/0407740 A1 CUI et al. ( 43 ) Pub . Date : Dec. 31 , 2020 ( 54 ) MATERIALS AND METHODS FOR Publication Classification CONTROLLING BUNDLE SHEATH CELL ( 51 ) Int. CI . FATE AND FUNCTION IN PLANTS C12N 15/82 ( 2006.01 ) ( 71 ) Applicant: FLORIDA STATE UNIVERSITY ( 52 ) U.S. CI . RESEARCH FOUNDATION , INC . , CPC C12N 15/8225 ( 2013.01 ) ; C12N 15/8269 Tallahassee, FL ( US ) ( 2013.01 ) ; C12N 15/8261 ( 2013.01 ) ( 57 ) ABSTRACT ( 72 ) Inventors : HONGCHANG CUI , The subject invention concerns materials and methods for TALLAHASSEE , FL (US ); DANYU increasing and / or improving photosynthetic efficiency in KONG , BLACKSBURG , VA (US ); plants, and in particular, C3 plants. In particular, the subject YUELING HAO , TALLAHASSEE , FL invention provides for means to increase the number of ( US ) bundle sheath ( BS ) cells in plants , to improve the efficiency of photosynthesis in BS cells , and to increase channels between BS and mesophyll ( M ) cells . In one embodiment, a ( 21 ) Appl . No .: 17 / 007,043 method of the invention concerns altering the expression level or pattern of one or more of SHR , SCR , and / or SCL23 in a plant. The subject invention also pertains to genetically ( 22 ) Filed : Aug. 31 , 2020 modified plants , and in particular, C3 plants, that exhibit increased expression of one or more of SHR , SCR , and / or SCL23 . Transformed and transgenic plants are contemplated Related U.S. Application Data within the scope of the invention . The subject invention also ( 62 ) Division of application No. 14 / 898,046 , filed on Dec. concerns methods for increasing expression of photosyn 11 , 2015 , filed as application No. PCT US2014/ / thetically important genes in a plant, wherein one or more 041975 on Jun . 11 , 2014 . genes of interest are operably linked with a plant SHR , SCR ( 60 ) Provisional application No. 61 / 833,771 , filed on Jun . or SCL23 promoter sequence and expressed in a plant. 11 , 2013 . Specification includes a Sequence Listing . Patent Application Publication Dec. 31 , 2020 Sheet 1 of 10 US 2020/0407740 A1 FIG . 1A FIG . 1C FIG . 1E BS BS 29 BS X BS p 14 p 22 : 4 um 40 + 10 um 22 + 5um FIG . 1A - 1 FIG . 1C - 1 FIG . 1E - 1 Patent Application Publication Dec. 31 , 2020 Sheet 2 of 10 US 2020/0407740 A1 FIG . 1B FIG . 1D FIG . 1F BS V BS X X 21+ 8 um 35 + 10 um 40 + 12 um FIG . 1B - 1 FIG . 1D - 1 FIG . 1F - 1 Patent Application Publication Dec. 31 , 2020 Sheet 3 of 10 US 2020/0407740 A1 FIG . 2A FIG . 2B FIG . 2C BS BS BS V X p < 1 ? FIG . 2D FIG . 2E FIG . 2F X X 13p H P FIG . 2G FIG . 2H Patent Application Publication Dec. 31 , 2020 Sheet 4 of 10 US 2020/0407740 A1 I QSCR E FIG.3C 0.37 3DFIG. € ou 0 A L 0 23-2SC/ FIG.3B FIG3E. scr1- 3AFIG. Patent Application Publication Dec. 31 , 2020 Sheet 5 of 10 US 2020/0407740 A1 23-2scScr-1 4BFIG. Col sc/23-2 FIG.4D WsSCP-1 GlucoseFructoseSucrose 5 3 2 1 ColShr-2 FIG.4A FIG.4C SM SCI1- 23-2SC/ scl23-2 Patent Application Publication Dec. 31 , 2020 Sheet 6 of 10 US 2020/0407740 A1 23-2sc!SCP1- FIG.4E Sc/23-2 Patent Application Publication Dec. 31 , 2020 Sheet 7 of 10 US 2020/0407740 A1 FIG . 5A 1 2 1218 bp D o o B 120.0 % 100.0 % 80.0 % 60.0 % 40.0 % 20.0 % 0.0 % sc /23 1 , sc123 1 , sc123 2 , sc.23 2 , # 1 # 2 # 1 # 2 FIG . 5B Patent Application Publication Dec. 31 , 2020 Sheet 8 of 10 US 2020/0407740 A1 FIG . 6A FIG . 6C A C 03:33 B D cao FIG . 6B FIG . 6D Patent Application Publication Dec. 31 , 2020 Sheet 9 of 10 US 2020/0407740 A1 a b FIG . 7A FIG . 7B Patent Application Publication Dec. 31 , 2020 Sheet 10 of 10 US 2020/0407740 A1 SCR targets 26 w 24 1877 SHR targets SCL23 targets Lists contain 2539 unique elements FIG . 8 US 2020/0407740 A1 Dec. 31 , 2020 1 MATERIALS AND METHODS FOR with the active transport system , CO2 is effectively concen CONTROLLING BUNDLE SHEATH CELL trated in the bundle sheath cells , which in turn leads to the FATE AND FUNCTION IN PLANTS repression of the oxygenase activity of RUBISCO . Last but not least , each bundle sheath cell layer is associated with a CROSS - REFERENCE TO RELATED central cylinder of vascular tissue , which provides water and APPLICATIONS inorganic nutrients, and is surrounded by a single layer of [ 0001] The present application is a divisional of U.S. mesophyll cells , a feature characteristic of C4 plants called application Ser. No. 14 / 898,046 , filed Dec. 11 , 2015 , which the Kranz anatomy ( Wang et al . ( 2011 ) ) . is the National Stage of International Application Number [ 0005 ] Attempts to increase yield by expressing PEP car PCT /US2014 /041975 , filed Jun . 11 , 2014 , which claims the boxylase in both mesophyll and bundle sheath cells rice benefit ofU.S. Provisional Application No. 61 / 833,771 , filed have failed ( Taniguchi et al . ( 2008 ) ) , suggesting that the Jun . 11 , 2013 , each of which is hereby incorporated by mesophyll and bundle sheath cells must be engineered reference herein in its entirety, including any figures, tables , separately ( Kajala et al . ( 2011 ) ) . nucleic acid sequences , amino acid sequences , or drawings . [ 0006 ] Despite the pivotal role of bundle sheath cells in C4 photosynthesis, the mechanisms that determine their cell identity and patterning are still unknown. Extensive mutant SEQUENCE LISTING screening efforts in the past decades have identified several [ 0002 ] The Sequence Listing for this application is labeled maize and Arabidopsis mutants defective in chloroplast “ 2QK2480.TXT" which was created on Sep. 17 , 2020 and development in the bundle sheath cells ( Nelson ( 2011 ) ; is 187 KB . The entire contents of the sequence listing is Brutnell et al . ( 1999 ) ; Hall et al . ( 1998 ) ; Kinsman and Pyke incorporated herein by reference in its entirety. ( 1998 ) ; Petricka et al . ( 2008 ) ; Rossini et al . ( 2001 ) ) , but none of these mutants affects bundle sheath cell identity. BACKGROUND OF THE INVENTION [ 0007 ] Bundle sheath cells are a leaf cell type that forms [ 0003 ] With a rapidly growing world population and a single cell layer between the mesophyll cells and the dwindling natural resources , we are facing an enormous central vascular tissue . In C3 plants, both the mesophyll challenge of increasing crop yields while simultaneously cells and BS cells are photosynthetic, but the BS cells are improving the efficiency of resource utilization . In C3 small with fewer chloroplasts. In contrast, the BS cells are plants, a 3 - carbon molecule is the first product of carbon the major sites of photosynthesis in most C4 plants, whereas fixation , whereas in C4 plants, a 4 -carbon molecule is the the mesophyll cells are involved in CO2 fixation only . first product. Because C4 plants are much more efficient than Accordingly , the BS cells are much larger in size . The spatial C3 plants in photosynthesis as well as water and nitrogen separation of the two phases of photosynthesis into meso usage , particularly in hot climates ( Langdale ( 2011 ) ) , tre phyll and BS cells is one of the features that make C4 plants mendous efforts are being taken to introduce C4 photosyn significantly more efficient photosynthetically. Another fea thesis into economically important C3 crops ( Sage and Zhu ture that improves the photosynthetic efficiency in C4 plants ( 2011 ) ) , such as rice ( Hibberd et al . ( 2008 ) ) . It is estimated is the Kranz anatomy, characterized by an approximately 1 : 1 that yields can be increased by 50 % if rice is transformed ratio between mesophyll and BS cells . The close association into a C4 plant ( Hibberd et al . ( 2008 ) ) . between the two cell types facilitates metabolite transport, [ 0004 ] Evidence indicates that C4 plants have evolved which is critical for the C4 mechanism . In C3 plants , this multiple times from C3 plants ( Brown et al . ( 2011 ) ) , but all ratio is greater than 2 : 1 . C4 plants share some common features that make them [ 0008 ] Many important crops , such as rice ( Oryza sativa ) perform better . A critical innovation is the deployment of and wheat ( Triticum aestivum ) , are C3 plants. To meet the phosphoenolpyruvate ( PEP ) carboxylase as the enzyme for needs for food of a rapidly growing population, tremendous the initial fixation of CO2 in C4 plants. Unlike RUBISCO , efforts are being undertaken to introduce the C4 mechanism the enzyme used for the initial fixation of CO2 in C3 plants, into C3 crops ( Langdale, 2011 ) . For example , millions of PEP carboxylase does not have an oxygenase activity and dollars have been invested at the C4 rice consortium to therefore can maintain a high rate of photosynthesis even convert this important crop into a C4 plant ( von Caemmerer under conditions of low CO2 ( stomates partially closed ) and et al . , 2012 ) . Although the input is huge and the risk is high , high temperature (RUBISCO's oxygenase activity is stimu the potential reward is enormous. It is estimated that a 10 % lated at high temperatures ( Spreitzer et al . ( 2002 ) ) . Another increase in the photosynthetic efficiency would increase the critical feature of C4 plants is the separation of the two yield by 50 % ( Langdale , 2011 ) . However, to achieve C4 phases of photosynthesis , namely Co , fixation and carbo photosynthesis in C3 plants requires engineering of the BS hydrate biosynthesis, into the mesophyll and bundle sheath and mesophyll cells at many levels , including an increase in cells , respectively . To sustain a high rate of photosynthesis, the density of BS cells and modification of the physiology in C4 plants also have numerous plasmodesmata and various the BS and mesophyll cells .
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  • Proteome-Wide Characterization of Sugarbeet Seed Vigor and Its Tissue Specific Expression

    Proteome-Wide Characterization of Sugarbeet Seed Vigor and Its Tissue Specific Expression

    Proteome-wide characterization of sugarbeet seed vigor and its tissue specific expression Julie Catusse*†, Jean-Marc Strub†‡, Claudette Job*, Alain Van Dorsselaer†, and Dominique Job*§ *Centre National de la Recherche Scientifique-Universite´Claude Bernard Lyon 1, Institut National des Sciences Applique´es–Bayer CropScience Joint Laboratory, Unite´Mixte de Recherche 5240, Bayer CropScience, 14-20 rue Pierre Baizet, F69263 Lyon Cedex 9, France; and ‡Laboratoire de Spectrome´trie de Masse Bio-Organique, De´partement des Sciences Analytiques, Institut Pluridisciplinaire Hubert Curien, Unite´Mixte de Recherche 7178, Centre National de la Recherche Scientifique-Universite´Louis Pasteur, Ecole Europe´enne de Chimie, Mate´riaux et Polyme`res, 25 rue Becquerel, F67087 Strasbourg Cedex 2, France Edited by Roland Douce, Universite´de Grenoble, Grenoble, France, and approved April 11, 2008 (received for review January 19, 2008) Proteomic analysis of mature sugarbeet seeds led to the identifi- The use of metabolic inhibitors (␣-amanitin and cyclohexi- cation of 759 proteins and their specific tissue expression in root, mide) showed that transcription is not required for the comple- cotyledons, and perisperm. In particular, the proteome of the tion of germination in Arabidopsis, implying that the potential of perispermic storage tissue found in many seeds of the Caryophyl- germination is largely programmed during seed maturation on lales is described here. The data allowed us to reconstruct in detail the mother plant (4). Therefore, in this work, we have charac- the metabolism of the seeds toward recapitulating facets of seed terized sugarbeet seed¶ vigor by proteomics. This was challeng- development and provided insights into complex behaviors such as ing, however, because there are virtually no genomics data germination.