Enhancement of Yield,Grain Quality Characters, 2-Acetyl-1-Pyrroline Content,and Photosynthesis of Fragrant Rice Cultivars by Foliar Application of Paclobutrazol

Paclobutrazol is a well-known plant growth regulator. However, the application of paclobutrazol in fragrant rice production has not been reported. The present study conducted a field experiment with two cropping seasons and three fragrant rice cultivars to investigate the effects of paclobutrazol application on yield formation, grain quality, 2-acetyl-1-pyrroline (2-AP, key component of fragrant rice aroma) biosynthesis, and photosynthesis of fragrant rice. At the initial heading stage, paclobutrazol solutions at 0 (control), 30, 60, 100, and 120 mg L^?1 were foliar applied to fragrant rice plants, respectively. Compared with control, paclobutrazol treatments significantly (P?<?0.05) increased grain yield and seed-setting rate of fragrant rice cultivars by 6.77–22.82% and 5.09–25.66%, respectively. Increased contents of photosynthetic pigments (chlorophyll a, chlorophyll b, and carotenoid) and the improved net photosynthetic rate at the grain-filling stage were observed due to paclobutrazol treatments. The paclobutrazol application increased head rice rate and grain 2-AP content by 3.94–8.94% and 6.47–18.80%, respectively. Lower chalky rice rate and chalkiness were recorded in paclobutrazol treatment than in control. Moreover, foliar application of paclobutrazol increased proline and Δ1-pyrroline contents in fragrant rice by 7.90–32.12% and 13.21–34.90%. Overall, foliar application of paclobutrazol could enhance productivity, improve grain quality, and increase the 2-AP content of fragrant rice, and 100–120 mg L^?1 was considered the suggested concentration of paclobutrazol application in fragrant rice production.

     

Modulation of 2-Acetyl-1-pyrroline (2-AP) Accumulation,Yield Formation and Antioxidant Attributes in Fragrant Rice by Exogenous Methylglyoxal (MG) Application

Pot experiments were conducted to investigate the physiological and biochemical effect of exogenous MG application on two fragrant rice cultivars, Meixiangzhan and Yuxiangyouzhan. The MG application included four treatments: 1 mmol·L^–1 (MG1), 5 mmol·L^–1 (MG5), and 10 mmol·L^–1 (MG10), and deionized water as control (MG0). Results indicated that MG1 significantly increased the yield by 11.54–14.06% through promoting the seed-setting rate. Increases on grain 2-acetyl-1-pyrroline (2-AP) content at maturity were found in MG1. The contents of proline, Δ1-pyrrolin-5-carboxylic acid (P5C), pyrroline, and MG, and the activities of proline dehydrogenase (PDH), ornithine aminotransferase (OAT), and Δ1-pyrrolin-5-carboxylate synthase (P5CS) in plant tissues at all sampling stage were also increased in MG1 than MG0. In addition, although the malondialdehyde (MDA) content at all sampling stage was increased by MG application, MG1 increased the antioxidant enzyme activities and soluble protein content in all plant tissues and the pigment content in leaves. However, high-level MG (10 mmol·L^–1) application significantly decreased the yield and grain 2-AP content. Overall, this study suggested that exogenous MG application regulated the 2-AP accumulation, yield formation, and antioxidant attributes in fragrant rice, and that MG application at the concentration of 1 mmol·L^–1 increased the 2-AP content.

     

Enhancement of Ultrasonic Seed Treatment on Yield,Grain Quality Characters,and 2-Acetyl-1-Pyrroline Biosynthesis in Different Fragrant Rice Genotypes

Fragrant rice is popular for the good grain quality and special aroma. The present study conducted a field experiment to investigate the effects of ultrasonic seed treatment on grain yield, quality characters, physiological properties and aroma biosynthesis of different fragrant rice genotypes. The seeds of three fragrant rice genotypes were exposed to 1 min of ultrasonic vibration and then cultivated in paddy field. The results of present study showed that ultrasonic seed treatment increased grain yield of all fragrant rice genotypes but the responses of yield formation to ultrasonic were varied with different genotypes. Compared with control, ultrasonic seed treatment increased grain 2-acetyl-1-pyrroline (2-AP, the key component of fragrant rice aroma) content by 13.40%–44.88%. Ultrasonic seed treatment also reduced the crude protein contents in grains. The head rice rate, rice length, chalky rice rate, and chalkiness degree were influenced by ultrasonic for one or two fragrant rice genotypes. The activities of peroxidase and superoxide dismutase were also enhanced due to ultrasonic seed treatment. In conclusion, ultrasonic seed treatment increased grain, regulated grain aroma and quality, and improved stress resistance of fragrant rice varieties.     

Foliar Application of Brassinolide Induced Regulation of Grain Yield and Quality,Antioxidant Responses and Aroma in Fragrant Rice

Brassinolide (BR) is a new green plant growth regulator. The present field study was conducted on two fragrant rice cultivars (i.e., Meixiangzhan-2 and Xiangyaxiangzhan) to study the effects of foliar application of BR on fragrant rice performance. At the heading stage, BR solutions at 0.05, 0.15, 0.25 and 0.50 mg L-1 were sprayed on fragrant rice at 600 liters per hectare; these treatments were named as BR1, BR2, BR3 and BR4, respectively. The treatment sprayed with distilled water was taken as a control (CK). Compared with CK, the BR2 treatment significantly enhanced the chlorophyll concentration and net photosynthetic rate for Meixiangzhan-2, and Xiangyaxiangzhan. The BR3 and BR4 treatments increased the concentrations of chlorophyll a, chlorophyll b and carotenoid, and also enhanced net photosynthetic rate by 31.91% and 40.43%, respectively. Higher grain yields were recorded in the BR2 treatment than on CK on Meixiangzhan-2, while on Xiangyaxiangzhan, the BR3 and BR4 treatments increased grain yield compared with CK. In relation to CK, higher head rice rates were recorded in the BR2 treatment for Meixiangzhan-2, and in the BR3 and BR4 treatments for Xiangyaxiangzhan. BR treatments (BR2 for Meixiangzhan-2, BR3 and BR4 for Xiangyaxiangzhan) also significantly enhanced the activities of GPX, SOD and CAT by 10.22% to 23.00%, and reduced the malonaldehyde concentration. In addition, we observed that some BR treatments (BR2 for Meixiangzhan-2, BR3 and BR4 for Xiangyaxiangzhan) decreased the grain 2-acetyl-1-pyrroline concentration of fragrant rice.     

Production of aromatic three-line hybrid rice using novel alleles of BADH2

Aroma is a key grain quality trait that directly influences the market price of rice globally. Loss of function of betaine aldehyde dehydrogenase 2 (OsBADH2) affects the biosynthesis of 2-acetyl-1-pyrroline (2-AP), which is responsible for aroma in fragrant rice. The current study was aimed at creating new alleles of BADH2 using CRISPR/Cas9 gene editing technology under the genetic background of the japonica Ningjing 1 (NJ1) and indica Huang Huazhan (HHZ) varieties. Sensory evaluation and analysis using headspace solid-phase microextraction gas chromatography–mass spectrometry (HS-SPME-GC-MS) showed that the grains of the four homozygous T₁ lines with new alleles of BADH2 (nj1-cr ^BADH2-1, nj1-cr ^BADH2-2, hhz-cr ^BADH2-1 and hhz-cr ^BADH2-2) produced moderate fragrance and had significantly increased 2-AP content compared with wild-types. Moreover, there were no significant differences in the amylose content and gelatinization temperature among the four lines with new alleles of BADH2 to the wild-types. Thereafter, we crossed the HHZ background new alleles of BADH2 with CMS line Taonong 1A (TN1A) to produce a three-line hybrid variety B-Tao-You-Xiangzhan (BTYXZ) with increased grain aroma. The 2-AP content in grains of the improved BTYXZ-1 and BTYXZ-2 reached at 26.16 and 18.74 μg/kg, and the gel consistency of BTYXZ-1 and BTYXZ-2 increased significantly by 9.1% and 6.5%, respectively, compared with the wild-type Tao-You-Xiangzhan (TYXZ). However, the γ-aminobutyric acid (GABA) content in the improved three-line hybrid rice BTYXZ-1 (5.6 mg/100 g) and BTYXZ-2 (10.7 mg/100 g) was significantly lower than that of the TYXZ. These results demonstrated that CRISPR/Cas9 gene editing technology could be successfully utilized in improving aroma in non-fragrant japonica and indica varieties. In addition, the newly developed BADH2 alleles provided important genetic resources for grain aroma improvement in three-line hybrid rice.     

不同定植期和摘心方案下5个品种(系)茶用菊生长和产量性状的变化

为了比较5个茶用菊新品种(系)的产量水平,并筛选出获得高产的定植期和摘心方案,以 ‘苏菊10号’、‘苏菊12号’、‘苏菊13号’、‘CH1-44’、‘CH5-13’ 为试材,采用三因素裂区试验设计,主区为早、中、晚3个定植期,裂区为5个新品种(系),裂裂区为4种摘心方案,比较不同栽培措施下植株生长和产量的差异.结果表明: 5个新品种(系)中,‘CH5-13’和‘苏菊13号’产量相对较高,‘CH1-44’和‘苏菊10号’产量次之,‘苏菊12号’产量最低;5月27日中期定植、二次摘心措施下5个新品种(系)的株高、冠幅、单株花数、花径、单花鲜质量、单株产量和单位面积产量均显著优于其他处理,较5月7日和6月13日定植分别提高16.0%和19.0%、18.0%和22.8%、36.7%和42.2%、11.1%和2.3%、13.0%和4.0%、47.8%和36.6%、48.5%和36.7%.随着摘心时间的推迟,株高显著降低,二次摘心株高较不摘心降低50.2%;二次摘心处理的冠幅、单株花数、单花鲜质量、单株产量和单位面积产量最高,较不摘心依次提高17.0%、29.1%、5.5%、34.0%和34.8%.品种(系)、定植期、摘心方案3个因素对茶用菊生长性状和产量影响作用的大小依次为:定植期>品种>摘心.     

测墒补灌对不同穗型小麦品种耗水特性和干物质积累与分配的影响

在田间试验条件下, 以中穗型小麦(Triticum aestivum)品种‘山农15’和大穗型品种‘山农8355’为供试材料, 设置3个0-140 cm土层土壤相对含水量处理: W0 (拔节期65%, 开花期60%)、W1 (拔节期70%, 开花期70%)、W2 (拔节后8天70%, 开花后8天70%), 采用测墒补灌的方法补充土壤水分达到目标相对含水量, 对两个不同穗型小麦品种的耗水特性和干物质积累与分配进行了研究。结果表明: (1)两品种籽粒产量均以W0处理最低, ‘山农15’ W1和W2处理无显著差异, ‘山农8355’ W1处理显著高于W2处理; 两品种W1处理的水分利用效率和灌溉水利用效率均显著高于W2处理。‘山农15’ W1处理的籽粒产量和灌溉水利用效率分别显著低于和高于‘山农8355’的W1处理, 水分利用效率无显著差异; 两品种W2处理的籽粒产量、水分利用效率和灌溉水利用效率均无显著差异。(2)两品种总耗水量以W0处理最低, ‘山农15’ W1处理显著低于W2处理, ‘山农8355’两处理无显著差异; 两品种W1处理的土壤供水量及其占总耗水量的比例显著高于W2处理。‘山农15’ W1处理的总耗水量和灌水量占总耗水量的比例显著低于‘山农8355’, 土壤供水量占总耗水量的比例显著高于‘山农8355’; 两品种W2处理总耗水量, 土壤供水量及其占总耗水量的比例无显著差异。(3)两品种W1处理成熟期干物质积累量显著高于其他处理, W1处理提高了‘山农8355’开花后干物质积累量及其对籽粒的贡献率, 对‘山农15’无显著影响。‘山农15’ W1和W2处理成熟期干物质积累量显著低于‘山农8355’, 开花前贮藏同化物向籽粒的转运量和转运率、对籽粒的贡献率均显著高于‘山农8355’, 开花后干物质积累量及其对籽粒的贡献率低于‘山农8355’。综合考虑干物质积累与分配、籽粒产量、水分利用效率和灌溉水利用效率, W1处理是两品种节水高产的最佳土壤相对含水量处理。     

Pyrroline-5-Carboxylate Reductase in Soybean Nodules : Comparison of the Enzymes in Host Cytosol, Bradyrhizobium japonicum Bacteroids, and Cultures

Characteristics of pyrroline-5-carboxylate reductase (P5CR) from Bradyrhizobium japonicum bacteroids and cultured rhizobia were compared with those of the enzyme in soybean nodule host cytosol. Reductase from host cytosol differed from that in bacteroids in: (a) the effect of pH on enzymic activity, (b) the capacity to catalyze both reduction of pyrroline-5-carboxylic acid and NAD⁺-dependent proline oxidation, (c) apparent affinities for pyrroline-5-carboxylic acid, and (d) sensitivities to inhibition by NADP⁺ and proline. The K₁ for proline inhibition of P5CR in bacteroid cytosol was 1.8 millimolar. The properties of P5CR in B. japonicum and bacteroid cytosol were similar. The specific activities of P5CR in the cytosolic fractions of the nodule host and the bacteroid compartment were also comparable.     

Solubilization of a Proline Dehydrogenase from Maize (Zea mays L.) Mitochondria

L-Proline is oxidized to pyrroline-5-carboxylic acid in intact plant mitochondria by a proline dehydrogenase (EC 1.4.3) that is bound to the matrix side of the inner mitochondrial membrane (TE Elthon, CR Stewart [1981] Plant Physiol 67: 780-784). This investigation reports the first solubilization of the L-proline dehydrogenase (PDH) from plant mitochondria. The supernatant from NP-40-treated etiolated shoot mitochondria of maize, Zea mays L., reduced iodonitrotetrazolium violet in a proline dependent manner. The pH optimum for this activity was 8. The apparent K_m for proline was 6.6 millimolar. When supplied with proline, this solubilized PDH activity also synthesized pyrroline-5-carboxylic acid. The PDH activity was inhibited in vitro by 300 millimolar potassium chloride but not by 300 millimolar potassium acetate. The PDH activity had a molecular mass that was greater than 150 kilodaltons. Mitochondria were prepared from etiolated shoots grown in 100% water-saturated vermiculite (control) and 16% water-saturated vermiculite (stress). The specific activity of solubilized PDH from the stress treatment was 11% of the same activity from the control treatment. Oxygen uptake in the presence of proline and ADP (state 3 proline oxidation) by mitochondria from the stress treatment was 25% of the same rate by mitochondria from the control treatment. Mitochondria were also prepared 16 hours after rewatering the seedlings growing in the stress treatment. Both the solubilized PDH specific activity and state 3 proline oxidation returned to the control levels. The specific activities of the NAD⁺-dependent pyrroline-5-carboxylic acid dehydrogenase and cytochrome c oxidase in the solubilized preparations were unaffected by these stress and recovery treatments. Oxygen uptake rates by intact mitochondria in the presence of ADP and NADH, succinate or malate-pyruvate were also unaffected by these treatments.     

Purification of the putA gene product. A bifunctional membrane-bound protein from Salmonella typhimurium responsible for the two-step oxidation of proline to glutamate

In this paper we report the purification of a protein which is able to catalyze both the proline oxidase and the pyrroline-5-carboxylic acid dehydrogenase activities necessary for the oxidation of proline to glutamic acid. The purification involves the preparation of a crude membrane pellet, detergent solubilization, ammonium sulfate fractionation, and DEAE-chromatography. We are able to obtain an essentially pure preparation (greater than 95% pure) after only a 52-fold purification, demonstrating that the protein is a major protein in cells fully induced for proline utilization. Both proline oxidase and pyrroline-5-carboxylic acid dehydrogenase activities co-purity throughout our purification. Velocity sedimentation of the purified protein demonstrates that both proline oxidase and pyrroline-5-carboxylic acid dehydrogenase activities co-sediment. Early in the purification procedure we are able to detect two species of protein which have both proline oxidase and pyrroline-5-carboxylic acid dehydrogenase activities. Our procedure purifies only the larger molecular weight species. The purified protein is a dimer composed of identical 132,000-dalton subunits. Analysis of mutants defective for proline utilization demonstrate that the bifunctional enzyme is the putA gene product.     

A new method for the preparation of delta 1-pyrroline 5-carboxylic acid and proline.

Hydroxylysine was oxidized with sodium metaperiodate and the major product characterized as Δ¹-pyrroline 5-carboxylic acid by the visible spectrum of its o-aminobenzaldehyde complex and by its reduction to proline by subsequent treatment with sodium borohydride. The reduction product was positively identified as proline by proton nuclear magnetic resonance spectroscopy and mass spectral analysis. The pH dependence for the preparation of Δ¹-pyrroline 5-carboxylic acid was determined by a study of the yields of proline obtained by variation of the pH values of the oxidative step. These observations support the hypothesis of intramolecular cyclization of α-aminoglutaric γ-semialdehyde as the second step in the reaction mechanism.     

Mutant cell lines resistant to azetidine carboxylic acid: Quantitative and qualitative differences in pyrroline-5-carboxylate synthase activity

Mutant Chinese hamster lung fibroblasts were selected that are resistant to the proline analog L-azetidine-2-carboxylic acid. Resistance in the two mutant cell lines is associated with two distinct alterations in pyrroline-5-carboxylate synthase, the enzyme that catalyzes the proline biosynthetic step leading from glutamic acid to pyrroline-5-carboxylate. In one mutant cell line, pyrroline-5-carboxylate synthase specific activity is increased 30-fold over the level in control cells. In the other mutant line, pyrroline-5-carboxylate synthase activity is not increased, but the enzyme has become insensitive to inhibition by ornithine and proline.     

Effect of exogenous amino acids on the intracellular content of proline and other amino acids in Daucus carota cells

The effect of tryptophan on the biosynthesis of proline has been investigated. Cells of Daucus carota grown in B5 medium supplemented with 5×10^–4M tryptophan acquired the ability to grow in the presence of inhibitory concentrations of azetidine-2-carboxylic acid, an analog of proline. When trp was added to carrot cell cultures at sub-growth inhibiting concentrations, overproduction of intracellular free proline was observed. An increase was also observed for lys, his, ala, leu and phe. Likewise, the addition of asparagine, glutamic acid and phenylalanine to the medium stimulated the intracellular increase of free proline and other amino acids.Abbreviations A2CA azetidine-2-carboxylic acid - 2,4-D 2,4-dichlorophenoxyacetic acid - 5MT 5-methyltryptophan - P5C pyrroline-5-carboxylic acid - f.wt. fresh weight - d.wt. dry weight     

Season of grazing and stocking rate interactively affect fuel loads in Baikiaea plurijuga Harms woodland in northwestern Zimbabwe

Wildfire is a major disturbance in Baikiaea plurijuga Harms woodland savannas. We tested the hypothesis that the timing and intensity of herbivory influence fuel loads. We used three stocking rates namely light (three cows and four goats ha^?1), medium (six cows and eight goats ha^?1) and heavy (eleven cows and sixteen goats ha^?1) and three times of grazing namely early‐, middle‐ and late‐growing seasons. Season of grazing and stocking rate influenced herbaceous phytomass. Phytomass was generally the highest (53.5 g DM m^?2) in paddocks grazed during the early growing season and the lowest (27.8 g DM m^?2) in those grazed during the late growing season. Phytomass was also generally the highest (40.4 g DM m^?2) in lightly stocked paddocks and the lowest (32.7 g DM m^?2) in heavily stocked ones. Litter mass was the lowest (160.8 g DM m^?2) in paddocks grazed during the early season whereas there were no differences in ungrazed paddocks and those grazed during either mid‐ or late growing seasons (205.4 g DM m^?2). There was a negative relationship between litter mass and stocking rate. Baikiaea Benth. woodlands should be grazed during either the mid‐ or late‐growing season at stocking rates greater than 0.1 LU ha^?1 to reduce grass fuel loads.     

Characterization of fragrance in sorghum (<Emphasis Type="Italic">Sorghum bicolor</Emphasis> (L.) Moench) grain and development of a gene-based marker for selection in breeding

Fragrance is one of the most important and valued quality characters in sorghum and other foods and attracts a premium price in local and global trade. The allele of the SbBADH2 gene in fragrant sorghum cultivar E228 was characterized. A 1441 bp deletion extending from exon 13 to 15 was found rather than a deletion from exon 12 to 15 as had been reported earlier. This allowed the development and validation of a new perfect PCR-based marker for identification of fragrant sorghum accessions in breeding. The concentration of 2-acetyl-1-pyrroline (2AP) in the grain of this cultivar was estimated to be 6.5 ± 0.4 ppb using headspace solid-phase microextraction (HS-SPME) coupled with GC-MS. Flavor components of fragrant sorghum accession E228 (IC 568489) were analyzed and compared with the non-fragrant M35-1 cultivar. PCA analysis revealed that 2AP, benzothiazole, 2,3,5-trimethylpyridine, (1E)-1-ethylidene-1H-indene, cedrene, 2,4-bis(2-methyl-2-propanyl)phenol, 2-hexyl-1-octanol, and 2-butyl-1-octanol were among 25 compounds that were found in sorghum grain that may be contributing toward the aroma of fragrant sorghum. Proline and methylglyoxal contents were found to be higher in E228 than in M35-1, while SbBADH2 expression in E228 was half that in M35-1, suggesting a similar 2AP biosynthetic mechanism to that found in fragrant rice and soybean.     

川中丘陵区覆膜栽培再生稻对CH4排放的影响

为探讨覆膜栽培再生稻对CH_4排放的影响,采用静态箱-气相色谱法观测了川中丘陵区2016和2017年覆膜条件下再生稻田的CH_4排放通量。试验设置覆膜单季中稻(SR)和覆膜中稻-再生稻(SR-RR)两个处理。结果表明:SR-RR处理中稻季提前出现CH_4排放峰,再生季CH_4排放量少,约占两季总排放的8%—10%。全观测期内SR-RR处理两季的CH_4排放总量为103—306 kg/hm~2,比SR处理的单季排放量高11%—16%(P0.05)。SR-RR处理两季稻谷总产量为10.2—10.4 t/hm~2,比SR处理高出19%—22%(P0.05)。SR-RR处理单位产量的CH_4排放量为9.9—30.1 kg/t、,比SR处理减少6%(P0.05)。覆膜条件下种植再生稻,可保证水稻高产稳产,减少单位产量的CH_4排放量,值得推广。     

水稻和小麦根尖细胞壁多糖的铝积累能力比较

采用水培法比较4种禾本科植物水稻（Oryza sativa L.）、玉米（Zea mays L.）、高粱（Sorghum bicolor（L.）Moench）和小麦（Triticum aestivum L.）8个基因型的抗铝（Al）能力,并对他们在Al积累后细胞壁的多糖组分进行分析。结果显示,在5~200 μmol/L Al处理下,水稻抗Al能力较强,而小麦抗Al能力较弱。在50 μmol/L Al处理下,小麦根尖的果胶和半纤维素1含量的增幅明显高于水稻。水稻基因型‘日本晴’与‘浙辐802’的细胞壁Al含量分别占根尖总Al含量的78.7%和91.6%;小麦基因型‘扬麦18’与‘扬麦16’Al含量分别占根尖总Al含量的64.9%和72.1%。Al吸附-解吸实验结果显示,小麦根尖细胞壁上Al的吸附量高于水稻。研究结果表明,细胞壁是Al积累的主要部位,对Al敏感的水稻和小麦基因型细胞壁中的Al主要分布在果胶中;而对Al耐性较强的水稻和小麦基因型细胞壁中的Al主要分布在半纤维素1中。