Cell-Specific Expression of Rubisco Small Subunit and Rubisco Activase Genes in C3 and C4 Species of Atriplex

1992 ◽  
Vol 19 (1) ◽  
pp. 89 ◽  
Author(s):  
GS Hudson ◽  
RE Dengler ◽  
PW Hattersley ◽  
G Dengler
Keyword(s):  
In Situ Hybridisation ◽  
Rubisco Activase ◽  
Small Subunit ◽  
Bundle Sheath ◽  
Specific Expression ◽  
Bisphosphate Carboxylase ◽  
C4 Species ◽  
Rubisco Small Subunit ◽  
Rbcs Gene

In situ hybridisation techniques have been used to determine the distribution of mRNAs for ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco: EC 4.1.1.39) and Rubisco activase in leaves of Atriplex patula L. (C3) and A. rosea L. (C4). In A. patula, mRNA for Rubisco small subunit (encoded by the rbcS gene family) was found to accumulate in the mesophyll and bundle sheath, while in A. rosea it accumulated in the bundle sheath only, as shown previously for the C4 monocot Zea mays L. The spatial distribution of rca transcripts for Rubisco activase paralleled that for the rbcS transcripts in both C3 and C4 Atriplex species, providing evidence that Rubisco activase is required in cells only where Rubisco is present.

scholarly journals Detection of phosphoenolpyruvate and ribulose 1,5-bisphosphate carboxylase transcripts in maize leaves by in situ hybridization with sulfonated cDNA probes.

1989 ◽  
Vol 37 (4) ◽  
pp. 423-428 ◽  
Author(s):  
C Perrot-Rechenmann ◽  
M Joannes ◽  
D Squalli ◽  
P Lebacq
Keyword(s):  
In Situ Hybridization ◽  
Cell Types ◽  
Small Subunit ◽  
Bundle Sheath ◽  
Differential Distribution ◽  
Mesophyll Cells ◽  
Bisphosphate Carboxylase ◽  
Maize Leaves ◽  
Specific Mrnas

This report outlines an efficient in situ hybridization method for locating specific mRNAs in tissue cryosections using sulfonated cDNA probes. The method involves chemical modification of DNA probes by insertion of a sulfone radical on cytosine residues, which generates a specific epitope. Sulfonated DNA is then detected by using indirect immunochemical procedure. Alternatively, antibodies conjugated to fluorescein or to alkaline phosphatase were used for mRNA detection. In situ hybridization was developed to study aspects of mesophyll and bundle sheath cell differentiation in maize leaves. Our results indicate that phosphoenolpyruvate carboxylase (PEP C) mRNA is restricted to mesophyll cells, and the nucleus-encoded mRNA of the small subunit (SSU) ribulose 1,5-bisphosphate carboxylase (RuBP C) is limited to the cytosol of bundle sheath cells. Thus, using in situ hybridization, we have demonstrated that the differential distribution of PEP C and RuBP C proteins in the two cell types also reflects the location of their mRNAs. These data imply either a tissue-specific transcriptional regulation or a selective mRNA degradation.

Structure and Expression of the Euglena rbcS Gene Coding for the Small Subunit of Ribulose 1,5 Bisphosphate Carboxylase/Oxygenase

1992 ◽  
pp. 641-644
Author(s):  
Weil Jacques-Henry ◽  
Chan Raquel Lia ◽  
Keller Mario ◽  
Tessier Luc-Henri ◽  
Imbault Patrice
Keyword(s):  
Small Subunit ◽  
Bisphosphate Carboxylase ◽  
Gene Coding ◽  
Rbcs Gene

Rubisco production in maize mesophyll cells through ectopic expression of subunits and chaperones

2021 ◽  
Author(s):  
Amber M Hotto ◽  
Coralie Salesse-Smith ◽  
Myat Lin ◽  
Florian A Busch ◽  
Isabelle Simpson ◽  
...  
Keyword(s):  
Large Subunit ◽  
Ectopic Expression ◽  
Carbon Assimilation ◽  
Rubisco Activase ◽  
Small Subunit ◽  
Bundle Sheath ◽  
Subunit Gene ◽  
In Planta ◽  
Mesophyll Cells ◽  
Assembly Factors

Abstract C4 plants, such as maize, strictly compartmentalize Rubisco to bundle sheath chloroplasts. The molecular basis for the restriction of Rubisco from the more abundant mesophyll chloroplasts is not fully understood. Mesophyll chloroplasts transcribe the Rubisco large subunit gene, and when normally quiescent transcription of the nuclear Rubisco small subunit gene family is overcome by ectopic expression, mesophyll chloroplasts still do not accumulate measurable Rubisco. Here we show that a combination of five ubiquitin promoter-driven nuclear transgenes expressed in maize leads to mesophyll accumulation of assembled Rubisco. These encode the Rubisco large and small subunits, Rubisco Assembly Factors 1 and 2, and the assembly factor Bundle Sheath Defective 2. In these plants Rubisco large subunit accumulates in mesophyll cells, and appears to be assembled into holoenzyme capable of binding the substrate analog CABP. Isotope discrimination assays suggest, however, that mesophyll Rubisco is not participating in carbon assimilation in these plants, most likely due to a lack of the substrate ribulose 1,5-bisphosphate and/or Rubisco activase. Overall, this work defines a minimal set of Rubisco assembly factors in planta and may help lead to methods of regulating the C4 pathway.

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