Grain length (size) and weight are essential components of crop yield. To date, many QTLs/genes for these traits have been identified. GS3 encodes a putative transmembrane protein and functions as a negative regulator, and its larger-grain allele contains a nonsense mutation causing a 178-aa truncation (Fan et al., 2006). GL3.1/qGL3 encodes a putative protein phosphatase and also acts as a negative regulator of grain size (Qi et al., 2012;Zhang et al., 2012). Another negative regulator of grain size and weight is TGW6, which hydrolyzes indole-3-acetic acid (IAA)-glucose into IAA and glucose (Ishimaru et al., 2013). In contrast, GW6a is a positive regulator of grain weight, which encodes a novel histone H4 acetyltransferase (Song et al., 2015). Copy number variation at the GL7/GW7 locus causes elevated expression of GL7 and thus an increase in grain length (Wang et al., 2015a(Wang et al., , 2015b. GL2/GS2 encodes the plant-specific transcription factor OsGRF4, and its larger-grain allele harbors a mutation preventing cleavage by miR396c, resulting in elevated GL2/GS2 expression (Hu et al., 2015;Che et al., 2016). GLW7 encodes the plantspecific transcription factor OsSPL13, and high OsSPL13 expression is associated with larger grains (Si et al., 2016). These findings have greatly enhanced our understanding of grain length and weight regulation; however, there are still gaps in integrating these factors into genetic network(s). Here, we report a thorough dissection of the QTL composition of grain size and the characterization of a novel QTL, qTGW3, that regulates grain length and weight in rice.

The individual roles of three chloroplast CPN60 protomers (CPN60α, CPN60β1, and CPN60β2) and whether and how they are assembled into functional chaperonin complexes are investigated in Chlamydomonas reinhardtii. Protein complexes containing all three potential subunits were identified in Chlamydomonas, and their co-expression in Escherichia coli yielded a homogeneous population of oligomers containing all three subunits (CPN60αβ1β2), with a molecular weight consistent with a tetradecameric structure. While homo-oligomers of CPN60β could form, they were dramatically reduced when CPN60α was present and homo-oligomers of CPN60β2 were readily changed into hetero-oligomers in the presence of ATP and other protomers. ATP hydrolysis caused CPN60 oligomers to disassemble and drove the purified protomers to reconstitute oligomers in vitro, suggesting that the dynamic nature of CPN60 oligomers is dependent on ATP. Only hetero-oligomeric CPN60αβ1β2, containing CPN60α, CPN60β1, and CPN60β2 subunits in a 5:6:3 ratio, cooperated functionally with GroES. The combination of CPN60α and CPN60β subunits, but not the individual subunits alone, complemented GroEL function in E. coli with subunit recognition specificity. Down-regulation of the CPN60α subunit in Chlamydomonas resulted in a slow growth defect and an inability to grow autotrophically, indicating the essential role of CPN60α in vivo.

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BackgroundChloroplast chaperonin, consisting of multiple subunits, mediates folding of the highly abundant protein Rubisco with the assistance of co-chaperonins. ATP hydrolysis drives the chaperonin allosteric cycle to assist substrate folding and promotes disassembly of chloroplast chaperonin. The ways in which the subunits cooperate during this cycle remain unclear.ResultsHere, we report the first crystal structure of Chlamydomonas chloroplast chaperonin homo-oligomer (CPN60β1) at 3.8 Å, which shares structural topology with typical type I chaperonins but with looser compaction, and possesses a larger central cavity, less contact sites and an enlarged ATP binding pocket compared to GroEL. The overall structure of Cpn60 resembles the GroEL allosteric intermediate state. Moreover, two amino acid (aa) residues (G153, G154) conserved among Cpn60s are involved in ATPase activity regulated by co-chaperonins. Domain swapping analysis revealed that the monomeric state of CPN60α is controlled by its equatorial domain. Furthermore, the C-terminal segment (aa 484–547) of CPN60β influenced oligomer disassembly and allosteric rearrangement driven by ATP hydrolysis. The entire equatorial domain and at least one part of the intermediate domain from CPN60α are indispensable for functional cooperation with CPN60β1, and this functional cooperation is strictly dependent on a conserved aa residue (E461) in the CPN60α subunit.ConclusionsThe first crystal structure of Chlamydomonas chloroplast chaperonin homo-oligomer (CPN60β1) is reported. The equatorial domain maintained the monomeric state of CPN60α and the C-terminus of CPN60β affected oligomer disassembly driven by ATP. The cooperative roles of CPN60 subunits were also established.Electronic supplementary materialThe online version of this article (doi:10.1186/s12915-016-0251-8) contains supplementary material, which is available to authorized users.

The specific cochaperonin, chloroplast chaperonin (Cpn)20, consisting of two tandem GroES-like domains, is present abundantly in plant and algal chloroplasts, in addition to Cpn10, which is similar in size to GroES. How Cpn20 oligomers, containing six or eight 10-kDa domains, cooperate with the heptameric ring of chaperonin at the same time as encountering symmetry mismatch is unclear. In the present study, we characterized the functional cooperation of cochaperonins, including two plastidic Cpn20 homo-oligomers from Arabidopsis (AtCpn20) and Chlamydomonas (CrCPN20), and one algal CrCPNs hetero-oligomer, consisting of three cochaperonins, CrCPN11, CrCPN20 and CrCPN23, with two chaperonins, Escherichia coli GroEL and Chlamydomonas CrCPN60. AtCpn20 and CrCPNs were functional for assisting both chaperonins in folding model substrates ribulose bisphosphate carboxylase oxygenase from Rhodospirillum rubrum (RrRubisco) in vitro and complementing GroES function in E. coli. CrCPN20 cooperated only with CrCPN60 (and not GroEL) to refold RrRubisco in vitro and showed differential complementation with the two chaperonins in E. coli. Cochaperonin concatamers, consisting of six to eight covalently linked 10-kDa domains, were functionally similar to their respective native forms. Our results indicate that symmetrical match between chaperonin and cochaperonin is not an absolute requisite for functional cooperation.

For grain crops such as rice (Oryza sativa), grain size substantially affects yield. The histone acetyltransferase GRAIN WEIGHT 6a (GW6a) determines grain size and yield in rice. However, the gene regulatory network underlying GW6a-mediated regulation of grain size has remained elusive. In this study, we show that GW6a interacts with HOMOLOGUE OF DA1 ON RICE CHROMOSOME 3 (HDR3), a ubiquitin-interacting motif-containing ubiquitin receptor. Transgenic rice plants over-expressing HDR3 produced larger grains, whereas HDR3 knockout lines produce smaller grains compared to the control. Cytological data suggest that HDR3 modulates grain size in a similar manner to GW6a, by altering cell proliferation in spikelet hulls. Mechanistically, HDR3 physically interacts with and stabilizes GW6a in a ubiquitin-dependent manner, delaying protein degradation by the 26S proteasome. The delay in GW6a degradation results in dramatic enhancement of the local acetylation of H3 and H4 histones. Furthermore, RNA sequencing analysis and chromatin immunoprecipitation assays reveal that HDR3 and GW6a bind to the promoters of and modulate a common set of downstream genes. In addition, genetic analysis demonstrates that HDR3 functions in the same genetic pathway as GW6a to regulate grain size. Therefore, we identified the grain-size regulatory module HDR3–GW6a as a potential target for crop yield improvement.

A hybrid Genetic Algorithm (GA) and Levenberg–Marquardt (GA–LM) method is proposed for cell suspension measurement with electrical impedance spectroscopy. This algorithm combines the GA with global search ability and Levenberg–Marquardt (LM) algorithm with local search ability, which has the advantages of high accuracy and high robustness. First, GA–LM is compared with GA and LM algorithm separately by ideal simulation. Second, Gaussian noise is added to the ideal simulation data. The anti-noise ability of the GA–LM is discussed. Finally, experiments are conducted to verify the practicability of the proposed GA–LM method. In the experiment, GA–LM is used to fit the impedance spectrum of yeast suspensions with different volume fractions and active states. The results show that the GA–LM algorithm can converge to the real value that is set in the simulation under ideal numerical simulation conditions. In the simulation within 2% noise level, the mean relative error of the parameter solution is less than 4%, and the root mean square error of the fitting is less than 0.4. This method also performs well in fitting of the experimental data. In addition, the electric double layer resistance and cell membrane capacitance are selected as the main indicators for the identification of yeast suspension concentration and activity, respectively.

Two new tellurite halides, CdPb2Te3O8Cl2 and Cd13Pb8Te14O42Cl14 with mixed cationic layered structures, have been synthesized by high-temperature solution method. CdPb2Te3O8Cl2 crystallizes in the noncentrosymmetric Aba2 space group, built by [CdPb2Te3O8]...