Changquan Zhang

The WRKY transcription factor OsWRKY78 regulates stem elongation and seed development in rice

WRKY proteins are a large super family of transcriptional regulators primarily involved in various plant physiological programs. In present study, the expression profile and putative function of the WRKY transcriptional factor, WRKY78, in rice were identified. Real-time RT-PCR analysis showed that OsWRKY78 transcript was most abundant in elongating stems though its expression was detected in all the tested organs. The expression profiles were further confirmed by using promoter-GUS analysis in transgenic rice. OsWRKY78::GFP fusion gene transient expression analysis demonstrated that OsWRKY78 targeted to the nuclei of onion epidermal cell. Furthermore, OsWRKY78 RNAi and overexpression transgenic rice lines were generated. Transgenic plants with OsWRKY78 overexpression exhibited a phenotype identical to the wild type, whereas inhibition of OsWRKY78 expression resulted in a semi-dwarf and small kernel phenotype due to reduced cell length in transgenic plants. In addition, a T-DNA insertion mutant line oswrky78 was identified and a phenotype similar to that of RNAi plants was also observed. Grain quality analysis data showed no significant differences, with the exception of minor changes in endosperm starch crystal structure in RNAi plants. Taken together, these results suggest that OsWRKY78 may acts as a stem elongation and seed development regulator in rice.

Structural Changes of High-Amylose Rice Starch Residues following in Vitro and in Vivo Digestion

High-amylose cereal starch has a great benefit on human health through its resistant starch content. In this paper, starches were isolated from mature grains of high-amylose transgenic rice line (TRS) and its wild-type rice cultivar Te-qing (TQ) and digested in vitro and in vivo. The structural changes of digestive starch residues were characterized using DSC, XRD, (13)C CP/MAS NMR, and ATR-FTIR. TQ starch was very susceptible to digestion; its residues following in vitro and in vivo digestion showed similar structural characteristics with TQ control starch, which suggested that both amorphous and crystalline structures were simultaneously digested. Both amorphous and the long-range order structures were also simultaneously hydrolyzed in TRS starch, but the short-range order (double helix) structure in the external region of TRS starch granule increased with increasing digestion time. The A-type polymorph of TRS C-type starch was hydrolyzed more rapidly than the B-type polymorph. These results suggested that B-type crystallinity and short-range order structure in the external region of starch granule made TRS starch resistant to digestion.

Structural and functional properties of alkali-treated high-amylose rice starch

Native starches were isolated from mature grains of high-amylose transgenic rice TRS and its wild-type rice TQ and treated with 0.1% and 0.4% NaOH for 7 and 14 days at 35 °C. Alkali-treated starches were characterised for structural and functional properties using various physical methods. The 0.1% NaOH treatment had no significant effect on structural and functional properties of starches except that it markedly increased the hydrolysis of starch by amylolytic enzymes. The 0.4% NaOH treatment resulted in some changes in structural and functional properties of starches. The alkali treatment affected granule morphology and decreased the electron density between crystalline and amorphous lamellae of starch. The effect of alkali on the crystalline structure including long- and short-range ordered structure was not pronounced. Compared with control starch, alkali-treated TRS starches had lower amylose content, higher onset and peak gelatinisation temperatures, and faster hydrolysis of starch by HCl and amylolytic enzymes.

Development and high-throughput genotyping of substitution lines carring the chromosome segments of indica 9311 in the background of japonica Nipponbare.

Chromosome segment substitution lines (CSSLs) are useful for the precise mapping of quantitative trait loci (QTLs) and dissection of the genetic basis of complex traits. In this study, two whole-genome sequenced rice cultivars, the japonica Nipponbare and indica 9311 were used as recipient and donor, respectively. A population with 57 CSSLs was developed after crossing and back-crossing assisted by molecular markers, and genotypes were identified using a high-throughput resequencing strategy. Detailed graphical genotypes of 38 lines were constructed based on resequencing data. These CSSLs had a total of 95 substituted segments derived from indica 9311, with an average of about 2.5 segments per CSSL and eight segments per chromosome, and covered about 87.4% of the rice whole genome. A multiple linear regression QTL analysis mapped four QTLs for 1000-grain weight. The largest-effect QTL was located in a region on chromosome 5 that contained a cloned major QTL GW5/qSW5 for grain size in rice. These CSSLs with a background of Nipponbare may provide powerful tools for future whole-genome discovery and functional study of essential genes/QTLs in rice, and offer ideal materials and foundations for japonica breeding.

A maize phytochrome-interacting factor 3 improves drought and salt stress tolerance in rice

Abstract Phytochrome-interacting factor 3 (PIF3) activates light-responsive transcriptional network genes in coordination with the circadian clock and plant hormones to modulate plant growth and development. However, little is known of the roles PIF3 plays in the responses to abiotic stresses. In this study, the cloning and functional characterization of the ZmPIF3 gene encoding a maize PIF3 protein is reported. Subcellular localization revealed the presence of ZmPIF3 in the cell nucleus. Expression patterns revealed that ZmPIF3 is expressed strongly in leaves. This expression responds to polyethylene glycol, NaCl stress, and abscisic acid application, but not to cold stress. ZmPIF3 under the control of the ubiquitin promoter was introduced into rice. No difference in growth and development between ZmPIF3 transgenic and wild-type plants was observed under normal growth conditions. However, ZmPIF3 transgenic plants were more tolerant to dehydration and salt stresses. ZmPIF3 transgenic plants had increased relative water content, chlorophyll content, and chlorophyll fluorescence, as well as significantly enhanced cell membrane stability under stress conditions. The over-expression of ZmPIF3 increased the expression of stress-responsive genes, such as Rab16D, DREB2A, OSE2, PP2C, Rab21, BZ8 and P5CS, as detected by real-time PCR analysis. Taken together, these results improve our understanding of the role ZmPIF3 plays in abiotic stresses signaling pathways; our findings also indicate that ZmPIF3 regulates the plant response to drought and salt stresses.

Toward underlying reasons for rice starches having low viscosity and high amylose: physiochemical and structural characteristics.

BACKGROUND To understand the reasons for low starch viscosity in rice variety Q11 (Qing-lu-zhan 11), the physiochemical and structural characteristics of flours and starches were investigated and compared with another rice cultivar with similar high amylose but normal viscosity. RESULTS Our results showed that residual α-amylase activity and proteins were not the major causes of low starch viscosity in Q11 rice. Homogeneous small granule size and lower swelling power of high-amylose Q11 rice starches was one reason for the low swelling volume and thus the low pasting property. Q11 starch paste contained some partially swollen granules, which could increase the fluidity and thus cause the low paste viscosity. The small gelatinization enthalpy might be due to the lower crystallinity in Q11 starches. Moreover, Q11 starches consisted of more amylose with short chains, but also amylopectin with fewer short chains (DP 11-21) and more long chains (DP 22-54), which might be other important factors contributing to the low viscosity of Q11 starches. CONCLUSION These data can add to our understanding of the relationships between low viscosity and physiochemical properties, and will be helpful in elucidating the underlying mechanism of formation of low starch viscosity, as well as applications for low-viscosity rice starches.

Characterization of Grain Quality and Starch Fine Structure of Two Japonica Rice (Oryza Sativa) Cultivars with Good Sensory Properties at Different Temperatures during the Filling Stage

Temperature during the growing season is a critical factor affecting grain quality. High temperatures at grain filling affect kernel development, resulting in reduced yield, increased chalkiness, reduced amylose content, and poor milling quality. Here, we investigated the grain quality and starch structure of two japonica rice cultivars with good sensory properties grown at different temperatures during the filling stage under natural field conditions. Compared to those grown under normal conditions, rice grains grown under hot conditions showed significantly reduced eating and cooking qualities, including a higher percentage of grains with chalkiness, lower protein and amylose contents, and higher pasting properties. Under hot conditions, rice starch contained reduced long-chain amylose (MW 10(7.1) to 10(7.4)) and significantly fewer short-chain amylopectin (DP 5-12) but more intermediate- (DP 13-34) and long- (DP 45-60) chain amylopectin than under normal conditions, as well as higher crystallinity and gelatinization properties.

Biofortification of rice with the essential amino acid lysine: molecular characterization, nutritional evaluation, and field performance

Highlight Marker-free transgenic rice that over-accumulates the essential amino acid lysine has been generated by combining lines with endosperm-specific and constitutive engineering of lysine, and has been evaluated in the field.

Effect of Simultaneous Inhibition of Starch Branching Enzymes I and IIb on the Crystalline Structure of Rice Starches with Different Amylose Contents

Mutating or inhibiting genes encoding starch branching enzymes (SBEs) can increase the amylose content (AC) of cereals. We analyzed endosperm starches from three rice cultivars with different ACs and from transgenic lines derived from them. The transgenic lines had simultaneously inhibited SBE I and IIb genes. Compared with the starch from their wild-type parents, the starch from transgenic lines showed significantly increased apparent ACs and lamella size and decreased relative crystallinity, double helix content, and lamellar peak scattering intensity, and altered short-range ordered structure in the external region. These changes were more prominent in the line derived from the high-AC cultivar than in those derived from waxy and low-AC cultivars. Inhibiting both SBE I and IIb changed the crystalline structure of starch from A-type to CA-type in lines derived from waxy and low-AC cultivars, and from A-type to C-type in that derived from the high-AC cultivar.

Morphology and structural properties of high-amylose rice starch residues hydrolysed by amyloglucosidase

High-amylose starches are attracting considerable attention because of their potential health benefits and industrial uses. Enzyme hydrolysis of starch is involved in many biological and industrial processes. In this paper, starches were isolated from high-amylose transgenic rice (TRS) and its wild type rice, Te-qing (TQ). The morphological and structural changes of starch residues following Aspergillus niger amyloglucosidase (AAG) hydrolysis were investigated. AAG hydrolysed TQ starch from the granule surface, and TRS starch from the granule interior. During AAG hydrolysis, the content of amorphous structure increased, the contents of ordered structure and single helix decreased, and gelatinisation enthalpy decreased in TQ and TRS starch residues. The A-type polymorph of TRS C-type starch was hydrolysed faster than the B-type polymorph. The short-range ordered structure and B-type polymorph in the peripheral region of the subgranule and the surrounding band of TRS starch increased the resistance of TRS starch to AAG hydrolysis.