Varietal Differences in Cell Wall β-(1→3),(1→4)-Glucan and Nonstructural Carbohydrate in Rice Stems during the Grain Filling Stage

Abstract

The contribution of cell wall components and nonstructural carbohydrate (NSC) to grain filling in rice (Oryza sativa L.) was clarified by investigating the differences in the dynamics of hemicellulose, sugar composition of hemicellulose, β-(1→3),(1→4)-glucan, and NSC among cultivars with different grain filling capacities. This investigation was performed using the stems of standard, high yield and low harvest index (HI) cultivars. Hemicellulose concentration in stems tended to decrease slightly during the grain filling stage. This decrease was attributed to a decrease in β-(1→3),(1→4)-glucan concentration, which was detected as a decrease in glucose composition of hemicellulose in the stems during the grain filling stage. The rate of decrease and decrease in the amount of β-(1→3),(1→4)-glucan in the stems differed among the cultivars. These were higher in high yield and high HI cultivars than in relatively low yield and low HI cultivars. Moreover, a positive correlation was observed between the rate of decrease in β-(1→3),(1→4)-glucan and NSC, indicating similarities in the dynamics of β-(1→3),(1→4)-glucan and NSC among the cultivars. When the top half of panicle was removed, β-(1→3),(1→4)-glucan and NSC concentrations in the culm and leaf sheath did not decrease during the grain filling stage. Therefore, the β-(1→3),(1→4)-glucan in stems might be one of the sources that supply substrate to panicle as well as NSC.     

High (1→3,1→4)-β-Glucan Barley Fractions in Bread Making and their Effects on Human Glycemic Response

Barley flour (BF) from whole naked grain and two (1→3,1→4)-β-glucan-enriched fractions, a sieved fraction (SF) and a water-extracted fraction (WF), were produced and mixed with bread wheat flour (BW), for bread-making quality evaluation. Bread was baked in a pilot plant and analysed for sensory properties, proximate composition and (1→3,1→4)-β-glucan content. Four kinds of bread were produced from different mixtures of barley ingredients and bread wheat flour: a 100% BW ((1→3,1→4)-β-glucan content: total, 0·1%; soluble, 0·1%); a 50% BW and 50% BF (2·4%; 2·0%); a 50% BW and 50% SF (4·2%; 2·8%); an 80% BW and 20% WF (6·3%; 5·7%). Eight adults were fed test meals of each of the four breads, containing the same amount (50 g) of available carbohydrate, and glycemic indices calculated from finger-prick capillary blood samples. A linear decrease in glycemic index was found for increasing (1→3,1→4)-β-glucan content. The 20% WF bread had a 28% lower glycemic index than the plain BW and also showed the best scores for sensory attributes. This research confirms the effectiveness of viscous (1→3,1→4)-β-glucan in reducing postprandial blood glucose levels, even in foods with a high glycemic index. The enrichment technique, and water extraction/freeze-drying technique, could enable the use of barley as a source of a high-value fibre for reducing the glycemic index of traditional wheat-based foods such as bread, without negatively affecting their sensory characteristics.     

Distribution of (1-3)(1-4)-β-d-glucans in kernels of selected cultivars of naked and hulled barley

The objective of this study was to determine the distribution of (1-3)(1-4)-β-d-glucans in the kernels of three high-glucan lines of naked barley (STH 4561, STH 4671 and STH 4676), a low-glucan line (STH 4572), and two low-glucan comparative cultivars (naked – Rastik, and hulled – Stratus). The content of (1-3)(1-4)-β-d-glucans assayed with the enzymatic method varied within the range from 4.04 to 5.71% dry matter basis. Microspectrophotometric analysis revealed significant differentiation in the structure of the kernels and in the distribution of (1-3)(1-4)-β-d-glucans; it also indicated a necessity of applying such processing methods as are appropriate to the distribution of functional components. The distribution of functional components in the high-glucan lines of naked barley makes them ideal for utilization both in the milling technology and in the production of grits and flakes.     

Tagging Blast Resistance Gene Pi 1 in Rice (Oryza sativa) Using Candidate Resistance Genes

An F3 population derived from C101LAC/CO39 containing 90 lines was analyzed for blast resistance with 48 candidate genes developed from resistance gene analogs (RGA) and suppression subtractive library. Genetic analysis confirmed that blast resistance of the population was controlled by a single gene Pi 1. One of the candidate genes, R10 was identified as associated with the blast resistance gene on the long arm of chromosome 11 and mapped using a DH population derived from Azucena/IR64. A pair of PCR based primers was designed based on the sequence of R10 for marker-aided selection of the blast resistance gene. The recombination frequency between Pi 1 and the marker was estimated as 1.28%. It suggested that strategy of employing candidate genes is useful for gene identification and mapping. A new RFLP marker and the corresponding PCR marker for tagging of Pi 1 were provided.     

Characterization of O-β-glucosidase Activity in Vanilla planifolia and the change during the Curing Crocess of Vanilla Beans

香草兰是最重要的食品香料之一,其主要香味成分为香兰素,主要存在于香草兰豆荚中.香兰素一般由成熟豆荚中的香兰素葡萄糖苷经过漫长的发酵过程转化而来.O-β-葡萄糖苷酶在发酵生香的过程中起着关键作用.研究结果表明:香草兰的根、茎、叶和豆荚中都具有O-β-糖苷酶活性,豆荚活性最高,叶片活性最低;来自不同器官的O-β-糖苷酶的底物特异性一致,都能降解香兰素糖苷、2-NPG、4-NPG,都不能降解n-OG及硫代葡萄糖苷;香草兰糖苷酶提取物在50～60℃处理1 min,对酶活性影响很小,70℃处理1 min,酶活性丧失约40％,说明香草兰糖苷酶对高温具有一定耐受性,杀青后剩余糖苷酶活性在漫长的发酵过程中能够满足酶促反应的需求.     

Differential Expression of Whitebacked Planthopper Resistance in the japonicaJindica Doubled Haploid Rice Population under Field Evaluation and Seedbox Screening Test

Whitebacked planthopper (WBPH) -resistance in a japonica / indica doubled haploid (DH) rice population established from a cross between WBPH-resistant japonica Chunjiang 06 and susceptible indica TN1, was comparatively evaluated through a field experiment based on the WBPH immigrant density and standardized seedbox screening test (SSST). All the susceptible DH lines in the field experiment behaved accordingly in SSST. However, 35 of resistant 66 lines (53%) in the field, were categorized to susceptible groups in SSST. Likewise, there were no significant differences in WBPH immigrant densities among 70 DH lines that were highly resistant to susceptible in SSST. The results revealed that SSST could not evaluate properly WBPH resistance in the DH lines. Four QTLs for WBPH-resistance phenotyped by the immigrant density were detected on chromosomes 2, 3, 4, and 11. Of them, the QTL on chromosome 4 was the most effective (LOD 21.8, variance 78%). Five QTLs associated with seedling mortality were mapped on chromosomes 2, 3, 4, 5 and 6. In addition to the QTL (LOD 10.5, variance 68%) on chromosome 4, there was another major QTL (LOD 12.7, variance 71%) located on chromosome 5, which was SSST-specific but might be irrespective of the WBPH resistance traits.     

Distribution of the Classification Traits in the F2 Progeny of Two Crosses of indica/japonica in Rice (Oryza sativa)

Comparison of seed setting rate, plant height and Cheng′s morphological traits were made between the F2 progeny of indica japonica hybrids, 02428/Pei′ai 64 and Taichung 65/Teqing. There existed resemblance normal distribution in plant height, and the distribution had no relationship with the seed setting rate and plant height in F1 progeny, suggesting that there were no gametic selection for the plant height in the F2 progeny of indica japonica hybrids. An average seed setting rate of the F2 was equal to the F1 populations in the recombination of 02428/Pei′ai 64 in which there was an excellent cross compatibility, but the remarkable increase in the seed setting rate from the F1 to F2 in Taichung 65/Teqing, indicating that gametic selection only took place in the indica japonica crosses with low cross compatibility while null or little in the hybrids with high compatibility. There were many intermediate types among the leaf pubescence, color of hull at heading, and length of the first and second panicle internode, and these Cheng′s morphological traits was distributed normally; there were also a little intermediate types in hull hairness and phenol reaction with bi peak distribution; while length to width ratio of grain was inclined to the japonica types obviously showing a skewed distribution in the F2 population. Although Cheng′s morphological traits in F2 populations had inconsistent distribution types, Cheng′s morphological traits index showed normal distribution, which suggested that intermediate types, i.e. indicalinous or japonicalinous, were primary in F2 population.     

Molecular weight and structure units of (1→3, 1→4)-β-glucans in dough and bread made from hull-less barley milling fractions

Milling fractions of hull-less barley, and dough and bread with hull-less barley flour (40%) and wheat flour (60%) were analysed in an investigation of how the properties of (1→3, 1→4)-β-glucan were affected by milling, dough formation and bread making. Calcofluor average molecular weight $({\overline{M}}_{\text{cf}})$ and molecular weight distribution and the cellotriosyl/cellotetraosyl ratio of the (1→3, 1→4)-β-glucan were determined. Four different hull-less barley samples were milled to produce straight-run white flours, shorts, bran and whole-meal flours. The molecular weight distributions were unimodal for all fractions, and the $({\overline{M}}_{\text{cf}})$ range was between 117×10⁴ and 188×10⁴. These parameters were similar for all barleys, although $({\overline{M}}_{\text{cf}})$ was somewhat lower in white flour and bran fractions and somewhat higher in shorts and whole-meal. The cellotriosyl/cellotetraosyl ratio (1.5–1.8) was also similar in all fractions. Doughs and breads were made to study how flour type (sifted or whole-meal barley flour), water content, yeast, mixing time and fermentation time affect (1→3, 1→4)-β-glucan. The molecular weight distribution of (1→3, 1→4)-β-glucan was polymodal with two or three populations for all doughs and breads, and the $({\overline{M}}_{\text{cf}})$ decreased with increasing mixing and fermentation time. These results indicated that (1→3, 1→4)-β-glucan was degraded by endogenous β-glucanases in the barley and/or wheat flour. The molecular weight was not significantly affected by bread-baking and other factors. After mixing and fermentation the cellotriosyl/cellotetraosyl ratio was about 1.7–1.8 and was thus not significantly different from that of the flour blends. Thus to retain high molecular weight (1→3, 1→4)-β-glucan, which is important for its cholesterol-lowering effect, it is thus important to keep the mixing and fermentation time as short as possible when baking hull-less barley bread.     

The combined use of hull-less barley flour and xylanase as a strategy for wheat/hull-less barley flour breads with increased arabinoxylan and (1→3,1→4)-β-D-glucan levels

Bread-making with a composite flour (CF) consisting of 60% wheat flour (WF) and 40% hull-less barley flour, increased the total and soluble (1→3,1→4)-β-D-glucan and total arabinoxylan (AX) contents of dough and bread samples, but decreased the specific bread loaf volume. A xylanase insensitive to inhibition by Triticum aestivum L. xylanase inhibitor (TAXI) and xylanase inhibiting protein (XIP), increased loaf volume by 8.8 and 20.1% for WF and CF breads, respectively. Xylanase addition not only markedly improved loaf volume of CF bread, but also increased the soluble AX content of the WF and CF dough and bread samples because of conversion of water-unextractable AX into soluble AX. The xylanase had no impact on the extractability and molecular weight of (1→3,1→4)-β-D-glucan, but (1→3,1→4)-β-D-glucan was degraded during bread-making probably because of endogenous β-glucanase activity. Taken together, the results clearly show that the combined use of hull-less barley flour and a xylanase active during bread making, lead to palatable breads with high total and soluble AX and (1→3,1→4)-β-D-glucan contents. The sum of total AX and (1→3,1→4)-β-D-glucan was 1.70% for WF bread and 3.06% for CF bread, while the sum of soluble AX and (1→3,1→4)-β-D-glucan was 0.49 and 1.41% for control WF and CF xylanase supplemented breads, respectively.     

Quantitative Trait Loci and Epistatic Analysis of Seed Anoxia Germinability in Rice (Oryza sativa)

Anoxia germinability (AG) of 35 rice varieties was evaluated under different temperature and water submergence conditions. The shoot (including coleoptile) length of seedlings germinating under 30℃, 0.2 m water submergence for 5 days could be used as an optimal criterion for the AG evaluation of all the varieties. Differences were observed among the AGs of 359 varieties from different regions and subspecies with the optimized method. Moreover, 81 recombinant inbred lines derived from a cross of Kinmaze (japonica)/DV85 (indica) were used to detect quantitative trait loci (QTLs) conferring AG. A total of five QTLs for AG in the recombinant inbred population were detected on chromosomes 1, 2, 5 and 7, respectively. Phenotypic variations explained by each QTL ranged from 10.5% to 19.6%. Based on the directions of the additive effects, the alleles at three loci qAG-1, qAG-2 and qAG-7 from Kinmaze increased AG, while alleles at loci qAG-5a and qAG-5b from DV85 increased AG. Meanwhile, three pairs of epistatic loci were found to be located on chromosomes 2, 3, 5 and 11 with significant effects ranging from 16.7% to 48.8%, and the highest one 48.78%, was detected between C563–X182 on chromosome 3 and R830–X208 on chromosome 5.     

【Short Report】Influence of the Improved System of Rice Intensification (SRI) on Rice Yield,Yield Components and Tillering Characteristics under Different Rice Establishment Methods

Abstract

The impacts of the system of rice intensification (SRI) and conventional management (CM) on grain yield, yield components and tillering capacity were examined under 4 rice establishment methods transplanting (TP), seedling casting (SC), mechanical transplanting (MT) and direct seeding (DS). SRI produced significantly higher grain yield than CM under TP and MT but not under DS or SC. DS and SC produced much higher seedling quality than TP or MT, suggesting that robust seedlings with vigorous roots weaken the positive effect of SRI on rice yield. SRI produced a higher tillering rate than CM, but did not affect ear-bearing tiller rate significantly. Moreover, the net photosynthetic rate of the recent fully expanded leaf at mid-tillering stage was significantly higher in SRI than in CM under MT and TP. The obtained results also indicated that SRI increased biomass accumulation before heading and improved utilization of photosynthates in the grain-filling stage.     

Identification and Fine Mapping of a Gene Related to Pale Green Leaf Phenotype near the Centromere Region in Rice （Oryza sativa）

A thermo-insensitive pale green leaf mutant (pgl2) was isolated from T-DNA inserted transgenic lines of rice (Oryza sativa L. subsp. japonica cv. Nipponbare). Genetic analysis indicated that the phenotype was caused by a recessive mutation in a single nuclear-encoded gene. To map the PGL2 gene, an F2 population was constructed by crossing the mutant with Longtefu (Oryza sativa L. subsp. indica). The PGL2 locus was roughly linked to SSR marker RM331 on chromosome 8. To finely map the gene, 14 new InDel markers were developed around the marker, and PGL2 was further mapped to a 2.37 Mb centromeric region. Analysis on chlorophyll contents of leaves showed that there was no obvious difference between the mutant and the wild type in total chlorophyll (Chl) content, while the ratio of Chl a / Chl b in the mutant was only about 1, which was distinctly lower than that in the wild type, suggesting that the PGL2 gene was related to the conversion between Chl a and Chl b. Moreover, the method of primer design around the centromeric region was discussed, which would provide insight into fine mapping of the functional genes in plant centromeres.     

Cloning and Bioinformatical Analysis of Phospholipase Dα1 Gene and Its Promoter in the Latex of Rubber Tree (Hevea brasiliensis Muell.Arg.)

植物磷脂酶PLDα1与伤害信号转导密切相关,是伤害诱导内源茉莉酸(Jasmonic acid,JA)生物合成的关键酶之一.橡胶树胶乳PLDα1基因(HbPLDα1)表达的研究将有助于揭示橡胶树乳管细胞JA信号转导及其调控橡胶生物合成的机制.在EST序列的基础上,通过RACE和Genome Walking方法分别克隆了橡胶树胶乳的HbPLDα1基因及其启动子序列.HbPLDα1基因的cDNA全长为2 870 bp,包含长度为2427 bp的完整开放阅读框(ORF),具有典型的植物PLDα蛋白保守功能域,与同属大戟科的蓖麻和麻风树的PLDα1基因亲缘关系最近.HbPLDα1基因启动子区域长为1 559 bp,除含有TATA box和CAAT box等基本顺式作用元件外,还存在JA和脱落酸等激素响应元件以及干旱胁迫等环境信号响应元件,这表明HbPLDα1基因的表达可能受激素和环境信号的调控,在橡胶树乳管细胞对激素和环境信号的响应过程中发挥重要作用.     

Administration of Lupinus albus Gamma Conglutin (Cγ) to n5 STZ Rats Augmented Ins-1 Gene Expression and Pancreatic Insulin Content

Several studies support the health-promoting benefits of lupins, particularly lupin proteins. It has been demonstrated that Lupinus albus gamma conglutin (Cγ) protein lowered blood glucose levels; thus, Cγ showed promise as a new anti-diabetic compound for type 2 diabetes (T2D) treatment. The aim of this study was to evaluate the effect of Cγ on Ins-1 gene expression and on pancreatic insulin content in streptozotocin-mediated diabetic rats. Cγ was isolated from Lupinus albus seeds. Its identification was confirmed with polyacrylamide gel electrophoresis under native and denaturing conditions. We used streptozotocin (STZ) to induce T2D on the 5th day of life of newborn male Wistar rats (n5-STZ). After 20 weeks post-induction, these animals (glycemia?>?200 mg/dL) were randomly assigned to three groups that received the following one-week treatments: vehicle, 0.90 %?w/v NaCl (n5 STZ-Ctrl); glibenclamide, 10 mg/kg (n5 STZ-Glib); or Cγ, 120 mg/kg (n5 STZ-Cγ). Glucose and insulin levels were measured before and after treatment. Ins-1 gene expression was quantified using real time polymerase chain reaction and the pancreatic insulin content was evaluated with immunohistochemistry. Post-treatment, the n5 STZ-Cγ and n5 STZ-Glib groups showed reductions in glucose, increments in serum insulin, and increases in Ins-1 gene expression and beta cell insulin content compared to the n5 STZ-Ctrl group. The results showed that Cγ had beneficial effects on Ins-1 gene expression and pancreatic insulin content. These biological effects of Cγ strengthen its promising potential as a nutraceutical and/or new agent for controlling hyperglycemia.