Table of Content

12 November 2011, Volume 37 Issue 11

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CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS

Distribution of Allelic Variation for Genes of Vernalization and Photoperiod among Wheat Cultivars from 23 Countries

YANG Fang-Ping, HAN Li-Meng, YAN Jun, JIA Xian-Chun, ZHANG Yong, QU Yan-Yang, WANG Zhong-Wei, HE Zhong-Hu

Acta Agron Sin. 2011, 37(11):  1917-1925.  doi:10.3724/SP.J.1006.2011.01917

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Molecular markers for vernalization genes Vrn-A1, Vrn-B1, Vrn-D1 and Vrn-B3 and photoperiod gene Ppd-D1 were used to detect the presence of these genes among 755 cultivars from 23 countries. Days to heading and physiological maturity of these cultivars were also recorded in Anyang, Henan Province, China to provide information for their utilization in Chinese wheat breeding program. Frequencies of Vrn-A1, Vrn-B1, Vrn-D1, and vrn-A1+vrn-B1+vrn-D1 were13.0%, 21.1%, 15.6%, and 64.2%, respectively. Dominant allele Vrn-B3 was absent in all tested materials. Dominant vernalization alleles Vrn-A1, Vrn-B1, and Vrn-D1 were mainly observed in Chinese spring wheat and middle and upper Yangtze Valley winter wheat regions, Italy, India, Japan, Canada, Mexico, Chile, Argentina, and Australia with spring type, while cultivars carryied all recessive alleles at the four vernalization loci. The gene recombination of vrn-A1, vrn-D1, and Vrn-B1 was found in winter wheat regions of Northern China, middle and southern US, Germany, France, Norway, Ukraine, Russia, Turkey, Iran, Hungary, Bulgaria, Romania, and Serbia, where the wheat growth habit is winter type.The frequency of Ppd-D1a was 55.2%, and photoperiod sensitive allele Ppd-D1b was mainly observed in cultivars from higher latitude regions of US, Germany, Norway, Hungary, Northeastern China, Canada, Chile and Argentina; while photoperiod insensitive allele Ppd-D1a was observed in the other wheat-growing regions. Most of cultivars with photoperiod insensitive allele Ppd-D1a could completephysiological maturity in Anyang, whereas cultivars from Germany, Norway, Hungary, Northwestern US, Northeast China, Chile and Argentina could not mature well. In Anyang, flowering time was not speeded up by the presence of dominant vernalization allele Vrn-A1a, cultivars with Vrn-B1 and Vrn-D1 could head normally due to the completion of vernalization requirement during winter season.

Cloning of Rhamnosyl Transferase Gene and Construction of Its RNAi Vector in Potato

WANG Wang-Tian, ZHANG Jin-Wen, WANG Di, ZHANG Jun-Lian, SI Fu-Jun, TAO Shi-Hang

Acta Agron Sin. 2011, 37(11):  1926-1934.  doi:10.3724/SP.J.1006.2011.01926

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Steroidal glycoalkaloids (SGAs) are potentially harmful metabolites found in potatoes and other Solanaceous plants. SGAs accumulation affects food quality and safety, we hope to use molecular biology methods to reduce SGAs content and assist breeding efforts to ensure food safety, develop new and improved varieties of potatoes. To investigate the relationship between the gene expression of the rhamnosyltransferase (sgt3) and the biosynthesis of steroidal glycoalkaloid (SGAs), and reveal the role of sgt3 gene in biosynthetsis of potato steroidal glycoalkaloid, similar gene cDNA sequence of the potato sgt3 was obtained from the total RNA of potato tubers by reverse transcription polymerase chain reaction (RT-PCR) using specific primers synthesized according to the conserved domain of GenBanK No: DQ266437 sequence. The protein function and structure of the similar sgt3 gene cDNA sequence were predicted and analyzed by related software of bioinformatics technology. Interference expression vector based on sgt3 gene was constructed. The Blast result showed that the gene sequence shared a high level of similarity with the sgt3 gene in GenBank (accession No: DQ266437) and its homology was 99.54%, and similarity of the amino acid sequence was 99%. The full-length of cDNA was 1 500 bp, which contained 505 amino acids, UDPG glycosyltransferase conserved domain and many important functional sites. The 3D structure of protein was predicted by homology comparative modeling in Swiss-Model, the results showed that the 3D structure of SGT3 was highly similar to that of the glycosyltransferase, so it was inferred that SGT3 should be a member of glycosyltransferase superfamily that has function of steroidal glycoalkaloid. Sgt3 similar gene obtained here was rhamnosyl transferase gene, and its sequence was submitted with GenBank No: HM188447. The RNA interference transformation expression vector in which sgt3 gene was regulated by Actin and CIPP promoters was constructed, which will lay a solid foundation for the synthesis of alkaloids glycosides, further research of metabolism and cultivation of transgenic potato varieties with low indican alkaloids character.

Genetic Analysis of Rice Mutant L-05261 with Panicle Apical Abortion Trait

GAO Su-Wei, ZHANG Ling, MAO Bi-Gang, WANG Jiu-Lin, CHENG Zhi-Jun, WAN Jian-Min

Acta Agron Sin. 2011, 37(11):  1935-1941.  doi:10.3724/SP.J.1006.2011.01935

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In rice, panicle apical abortion (PAA) has a detrimental effect on the final yield. Using one PAA mutant L-05261, we found that the occurrence of PAA is related to the excess accumulation of hydrogen peroxides (H₂O₂). The hybrid plants of four cross-combinations from two less PAA varieties of 9311, PA64, and two non-PAA varieties of Balilla and IRAT129 crossed to L-05261 showed PAA phenotypes, on their F₂ populations, the percentages of PAA showed a continuous distribution and a bias towards non-PAA parents. The ratio of PAA plants and non-PAA plants in the L-05261×IRAT129 F₂ fitted to 63:1, while the ratio in the BC₁F₁consisted of 7:1, corresponding to a genetic model involving three or more partial dominant genes. One hundred and eighty-two individuals from F₂ of L-05261×IRAT129 were employed for QTL analysis, and four QTLs designated as qPAA3, qPAA4, qPAA5, and qPAA8 respectively, were detected out.However, no significantinteraction was found among these QTLs. Totally, all four QTLs were able to explain 46.32% of observed phenotypic variation. The remaining phenotypic variation was likely caused by environment effect, which should be considered in the following study.

Establishment and Application of Multiplex PCR System Based on Molecular Markers of Glutenin Subunit Genes (Loci) Related to Strong-gluten in Wheat

LIANG Qiang, ZHANG Xiao-Ke, WEI Qian, WANG Xiao-Long, ZHANG Jing, SUN Dao-Jie, FU Xiao-Ji

Acta Agron Sin. 2011, 37(11):  1942-1948.  doi:10.3724/SP.J.1006.2011.01942

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Wheat strong-gluten quality is closely correlated with combinations of high-molecular-weight glutenin subunits (HMW-GS) and low-molecular-weight glutenin subunits (LMW-GS). The multiplex PCR system is a rapid and efficient approach to evaluate wheat germplasm and its quality in wheat. In this study, we developed a multiplex PCR system confering molecular markers on Ax1/Ax2^*, Dx5, Glu-A3d, Glu-B3i genes, and Glu-B3 locus and validated it with 12 cultivars with known subunit at each locus. This multiplex PCR system was proved to be effective and stable to amplify target bands for these genes (locus), and used to evaluate the glutenin subunit genes (locus) accociated with strong-gluten in 62 major cultivars in wheat prodution in Shaanxi Province, China. The results showed that the frequencies of genes Ax1/Ax2^*, Dx5, Glu-A3d, Glu-B3i, and locus Glu-B3 in the 62 cultivars were 56.5%, 9.6%, 33.9%, 1.6%, and 64.4%, respectively, whereas gene Bx7^OE was not found. Most of the cultivars carried two-gene (locus) combinations with the frequency of 48.3%, a few cultivars carried a single gene or locus (33.9%). The frequency of cultivars carrying three or four-gene (locus) combinations was 11.3%. The remaining cultivars (6.5%) were free of above elite gene (locus). Therefore, the frequency of combination of multiple strong-gluten subunits gene (locus) was low in cultivars from Shaanxi Province, which could be promoted through germplasm introduction and traditional breeding aided by molecular marker selection. The multiplex PCR system developed in this study may serve as a rapid and efficient method to select materials pyrimiding multiple genes (loci) associated with strong-gluten in wheat breeding for quality.

Cloning of nia Gene and Its Differential Expression Induced by Different Nitrogen Forms in Sugar Beet (Beta vulgaris L.)

DING Guang-Zhou, HOU Jing, CHEN Li, MA Feng-Ming, CHEN Lian-Jiang

Acta Agron Sin. 2011, 37(11):  1949-1955.  doi:10.3724/SP.J.1006.2011.01949

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From diploid species Ty7, a cDNA clone related to nitrate reduction was isolated by Homology-based cloning. According to its sequences information, a novel full-length cDNA termed nia (accession number: EU163265) was obtained by using rapid amplification of cDNA ends (RACE). Nia was 3 247 bp long containing a 2 718 bp ORF, encoded 905 amino acids with a theoretical molecular weight of 102 kD and an isoelectric point of 6.12. Southern bloting proved that nia gene of Ty7 existed in the form of low copies. It was also proved that this clone was the style of NADH-NR. To reveal the effect of nitrate and ammonium nitrogen on nia expression, we determined NR activity in sugar beet under the treatment of nitrate and ammonium. Gene transcripts of nia were detected by semi-quantitative PCR. Efficiency of mRNA synthesis from each sample was estimated by quantitative PCR of glyceraldehyde-3-phosphate dehydrogenase (GAPDH). The results showed that when nia gene was induced by ammonium, the gene expression was promoted with low concentration of ammonium ion, while inhibited by high concentration of ammonium ion. When nia gene was induced by nitrate, being positively correlated with the concentration, the gene expression was enhanced with the increase of the concentration. By 30 mmol L^–1 nitrate induction for four hours, nia gene expression reached the highest value, and at about six hours of induction, the expression decreased significantly.

Resistance of CIMMYT Wheat Germplasm to Heterodera filipjevi Xuchang Population from Henan Province, China

YUAN Hong-Xia, ZHANG Fu-Xia, ZHANG Jia-Jia, HOU Xin-Song, LI Hong-Jie, LI Hong-Lian

Acta Agron Sin. 2011, 37(11):  1956-1966.  doi:10.3724/SP.J.1006.2011.01956

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The cereal cyst nematode (CCN) of wheat has become a severe disease in recent years. Heterodera filipjevi is a new pathogenic nematode of CCN in China. For screening resistant resources in breeding to H. filipjevi, the resistance of 75 wheat germplasm cultivars or lines from CIMMYT was evaluated in the greenhouse and field using relative resistance index (RRI) and Pf/Pi ratios, with the nematode from Xuchang population, Henan Province, China. No cultivar was immune to H. filipjevi. However, line 6R(6D) was highly resistant in two test conditions. CROC_1/AE.SQUARROSA (224)//OPATA^*1, CROC_1/AE.SQUARROSA (224)//OPATA*2, MACKELLER, CPI 133842, CPI 133814, and TRIDENT were moderately resistant to H. filipjevi in greenhouse. In the field test, CPI 133842, CPI 133814, DURATI, and TURCAN#39 showed a high resistance with RRI values ranging from 0.90 to 0.96. Fourteen lines including ID-2150, BAXTER, and MACKELLER were moderately resistant to H. filipjevi. The disease severity was heavier in greenhouse than in the field, and resistance identification in the greenhouse is much easier and reliable. The results indicated that relative resistance index (RRI) is an efficient index for evaluating the resistance of wheat cultivars to CCN.

Identification of SSR Markers Linked to Oil Content in Peanut (Arachis hypogaea L.) through RIL Population and Natural Population

HUANG Li,ZHAO Xin-Yan,ZHANG Wen-Hua,FAN Zhi-Ming,REN Xiao-Ping,LIAO Bo-Shou,JIANG Hui-Fang,CHEN Yu-Ning

Acta Agron Sin. 2011, 37(11):  1967-1974.  doi:10.3724/SP.J.1006.2011.01967

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Oil content has been an important quality trait for peanut. However, the progress in genetic improvement of oil content is slow. This is mainly due to the complex genetic basis, the high cost in oil content testing, and difficulty in phenotypic selection because of environmental influence. Therefore, marker assisted selection (MAS) is necessary for oil content improvement in peanut breeding. In the present study, F₈ RILs derived from a cross of Yuanza 9102×Zhonghua 5 were selected as materials. Ten RILs with extremely low oil content (about 52.91% on an average), twelve RILs with extremely high oil content (about 58.85% on an average) and the two parental lines were used to screen 631 simple sequence repeat (SSR) primers. Thirteen polymorphic bands of seven SSR primers were able to distinguish the extremely high and low oil content lines. Subsequently, the seven SSR primers were used to genotype the total RIL population. Based on the statistics of the SSR data from 20 lines with low oil content (less than 55%) and 45 lines with high oil content (more than 56%), we identified that a SSR marker, 2A5-250/240, was tightly linked to the oil content trait. The band 2A5-250 appeared almost in all low oil content lines with the match rate of 95.0%, while the band 2A5-240 appeared almost in all high oil content lines with the match rate of 88.9%. Detection of the marker 2A5-250/240 was conducted in 11 high oil (55.93% on an average) and 11 low oil (48.41% on an average) content peanut cultivars. The results showed that the match rates of the band 2A5-250 to low oil content materials and 2A5-240 to high oil content materials was 90.9% and 63.6%, respectively. In addition, the band 2A5-240 appeared in 10 of the 19 wild peanut (Arachis) lines with high oil content (about 58.60%). The results based on the analyses using RIL and natural populations demonstrated that the SSR marker 2A5-250/240 was useful for marker-assisted selection of oil content in peanut.

CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS

Establishment of Molecular Identity in 142 Sweet Sorghum Varieties

WANG Li-Meng, JIAO Shao-Jie, JIANG Yan-Chi, YAN Hong-Dong, SU De-Feng, SUN An-Quan

Acta Agron Sin. 2011, 37(11):  1975-1983.  doi:10.3724/SP.J.1006.2011.01975

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A large change occurs in phenotypic characters under different environments for introducing and frequently exchanging of abundant varieties, and accurate identification is usually affected by the similar phenotypic characters performed in the field, so it is necessary to establish ID to distinguish varieties between each other. A total of 103 SSR primers covering 10 linkage groups of sorghum chromosome were screened, 41 primers showing polymorphic bands were used to establish molecular ID of 142 sweet sorghum germplasm resources. A total of 189 polymorphic bands were amplified and obtained. The number of alleles revealed by each primer was 2-11 with an average of 4.6 per primer. The polymorphism information content (PIC) value for the SSR loci ranged from 0.089 to 0.850 with an average of 0.543. There existed much difference in specific indices of varieties, ranging from 109.1 to 454.7 with an average of 189.0. The results showed that 11 primers (Xtxp329, Xtxp258, Xtxp113, Xtxp303, Xtxp61, Xtxp201, Xtxp141, Xtxp91, Xtxp47, Xtxp217, and Xtxp67) were confirmed on the basis of allele number of each pair of primers. The IDs of 142 sweet sorghum germplasm resources were established by the 11 pairs of primers, which can discriminate a variety from others.

Function and Expression Specificity Analysis of Arabidopsis thaliana Gene AT2G14260

WANG Fei-Fei, LI Yong, WANG Hua-Dong, ZHU Yan-Meng, CAI Hua, JI Wei, BAI Ti

Acta Agron Sin. 2011, 37(11):  1984-1990.  doi:10.3724/SP.J.1006.2011.01984

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The AT2G14260 gene encodes proline iminopeptidase in Arabidopsis thaliana. Genechip expression profile indicate AT2G14260 responses to abiotic stress, such as high salt or cold. To analyze the function and expression specificity, we compared the wild type with mutants pip-1 under cold, drought and salt stresses. Under normal conditions, mutant plants showed similar growth performance with wild type plants. However, after stress treatments, mutant plants exhibited much shorter roots compared with wild type plants. In addition, the leaves of mutant plants were notably more yellow than wild-type leaves under drought stress. The average relative proline contents in wild type plants were 1.11, 1.23, 1.10, and 1.34 times of those in mutants. All these results showed that AT2G14260 gene plays an important role in plant tolerance to abiotic stress. Meanwhile, we isolated the gene promoter and constructed plant vector in which the GUS gene was expressed by AT2G14260 promoter, and then transformated into Arabidopsis thaliana. The results of the GUS staining revealed that the GUS gene did not express under normal conditions. After treatments, GUS gene expressed in the roots, stems and leaves of the transgenic seedlings at the two-leaf stage and the blooming stage, especially in the petals and flowers under drought stress or in the stigma under cold stress. Results of GUS relative activity measurement suggested this gene had the response ability to stress. Taken together, the promoter of AT2G14260 is stress inducible and tissue specific. Thus, AT2G14260 gene and its promoter have application potentials in genetic engineering.

Transgenic Rice Lines Harboring McCHIT1 Gene from Balsam Pear (Momordica charantia L.) and Their Blast Resistance

ZHANG Chang-Wei, LING Yang-Hua, SANG Xian-Chun, LI Bing, ZHAO Fang-Meng, YANG Zheng-Lin, LI Yun-Feng, FANG Li-Kuai, HE Guang-Hua

Acta Agron Sin. 2011, 37(11):  1991-2000.  doi:10.3724/SP.J.1006.2011.01991

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Rice blast is one of the most important diseases damaging rice. It is important to explore a novel way of improving blast resistance with introducing exogenous resistance genes in modern breeding. In the study, identification and screening of blast resistance in the transgenic rice offspring were conducted using methods of both seedling nursery artificial inoculationand natural infection in field nursery under the conditions of higher resistance selection pressure increased gradually from T₂ generation. Seven stable and outstanding McCHIT1-transforming rice lines with the most significant blast resistance were obtained in T₅ generation. Detected with 115 Magnaporthe grisea effective isolates from 26 physiological races belonging to seven groups, the resistance frequenciesfor the seven lines were 52.2–61.4%, with 16 percentage higher than those (36.8%) for the control Jinhui 35. Compared with the control, these lines increased the resistance to ZE race group, and improved the resistance to the races of group ZG and ZF as well as dominant group ZB. The McCHIT1-transforming rice lines C36-2-1, C21-6-2, and C21-3-1 were the better combination of higher yield and resistance with seed setting rate of over 80%. The McCHIT1 gene is a certain broad-spectrum resistance gene. The disease-resistant lines with both broad-spectrum resistance and better yield traits can be obtained effectively by introducing McCHIT1 gene under gradually increased selection pressure and screening excellent resistance from resistant lines.

TILLAGE & CULTIVATION·PHYSIOLOGY & BIOCHEMISTRY

Effect of Lipid Content and Components on Cooking Quality and Their Response to Nitrogen in Milled Japonica Rice

GU Dan-Dan, LIU Zheng-Hui, LIU Yang, WANG Chao-Hua, WANG Jiang-Cheng, LI Gang-Hua, DING Yan-Feng

Acta Agron Sin. 2011, 37(11):  2001-2010.  doi:10.3724/SP.J.1006.2011.02001

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In present study, six japonica cultivars and seven nitrogen treatments were used to study the effect of lipids content and components in milled rice on amylose content and RVA profile characteristics and the relationship between them and nitrogen. The results showed that the content and components of lipid in milled rice were significantly affected by genotype. As compared with large panicle cultivars, small panicle cultivars had higher crude lipids and starch lipids and lower non-starch lipids in milled rice. As compared with genotype, nitrogen had little effect on the content and components of lipid in milled rice. The content and components of lipid in milled rice were notably related to amylose content and the RVA profile characteristics. The crude lipid content was significantly and negatively correlated to BDV (breakdown), and starch lipids content was significantly and negatively correlated to SBV (setback) and CSV (consistency viscosity). From these results we summarized that the lipid content especially the starch lipids content in milled rice has significant effect on the cooking and eating quality of rice, which will be promoted with the increase of starch lipids content in milled rice.

Population Quality of Rice under Different Irrigation Regimes

ZHANG Zi-Chang, XU Yun-Ji, CHU Guang, WANG Zhi-Qin, WANG Hua-Meng, LIU Li-Jun, YANG Jian-Chang

Acta Agron Sin. 2011, 37(11):  2011-2019.  doi:10.3724/SP.J.1006.2011.02011

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Establishment of a population with high quality is an important prerequisite to achieve high grain yield. Irrigation is one of the most important practices in rice production and plays a critical role in regulation of population quality. To understand the effect of irrigation regimes on population quality, we planted two rice cultivars currently used in the production, Yangdao 6 (an indica) and Yangjing 4038 (a japonica), treated with furrow irrigation between beds (FI), alternate wetting and soil drying (AWD), and traditional flooding as control (TF), for 10 days after transplanting to maturity. Compared with TF, FI and AWD increased grain yield by 10.60% and 8.43%, respectively. Both FI and AWD regimes reduced the number of unproductive tillers and increased percentage of productive tillers, area and length of top three leaves, grain-leaf area ratio, light transmission rate, dry matter accumulation from heading to maturity, and increased photosynthetic rate of flag leaf, root activity and root biomass during grain filling. The results demonstrate that both FI and AWD could improve population quality, and consequently, increase grain yield.

Effects of Soil Fertility and Nitrogen Application Rates on Super Rice Yield, Quality, and Nitrogen Use Efficiency

ZHANG Jun, ZHANG Hong-Cheng, DUAN Xiang-Mao, XU Zong-Jin, YANG Bei, GUO Bao-Wei, DU Bin, DAI Ji-Gen, HU Ke, HE Zhong-Xiang, WEI Hai-Yan

Acta Agron Sin. 2011, 37(11):  2020-2029.  doi:10.3724/SP.J.1006.2011.02020

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Super rice Xudao 3 was adopted as material, the effect of nitrogen application (0, 148. 5, 223.5, 297.0, 372.0, and 445.5 kg ha^-1) under different soil fertility levels on yield, yield components, nitrogen use efficiency and quality of super rice was studied. Results were as follows: (1) The yield of Xudao 3 showed the trend under the roil with high fertility>medium fertility >low fertility; and the corresponding optimal nitrogenapplication to the highest yield for the three soil fertility levels were 260.8, 290.5, and 345.5 kg ha^-1, respectively. (2) There were significant or highly significant quadratic relationships betweenapparent N recovery efficiency (ANRE) and nitrogen application; andthe corresponding optical application of nitrogen to the highest ANREunder high, medium and low soil fertilities were 268.6, 293.4, and 335.2 kg ha^-1, respectively. (3) Soil fertility optimization was able to improve rice’s nutritional quality, processing quality and cooking-eating quality. And apllying the optimal nitrogen to the soil with different fertility levels could improve the appearance quality of rice and optimize the nutritional quality of rice. Based on the above nitrogen application for high-yield, high quality, efficient nitrogen utilization, we suggested that the nitrogen application of super rice for the soil with high, medium and low soil fertilities were 240–270 kg ha^-1, 285–315 kg·ha^-1, and 330–360 kg ha^-1, respectively.

Contents of Protein Components Stored in Grains and Activities of Related Enzymes in Wheat Cultivars in Different Quality Types

SHI Yu, GU Chu-Bei, YU Zhen-Wen, HU Zhen-Zhu

Acta Agron Sin. 2011, 37(11):  2030-2038.  doi:10.3724/SP.J.1006.2011.02030

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Stored protein content and its components are determinative factors of wheat processing quality. The activities of enzymes involved in nitrogen metabolism impact the accumulations of protein components. However, the effect of enzyme activities on stored protein content in grain is not clearly understood. In this study, we used six wheat cultivars grouped into I-type (GC8901 and 9411, first-class strong gluten), II-type (Jinan 17 and Yannong 19, second-class strong gluten), and III-type (Taishan 23, and Lumai 21, medium gluten) to observe the dynamic accumulations of stored protein components in grains and the activities of related enzymes in leaves during grain filling. The contents of total protein and stored protein were not significantly different between I-type and II-type, but type-I had higher glutenin content and lower ratio of Gli-to-Glu than type-II. Gliadin contents were presented with the order of type-II > type-I > type-III from 20 d after anthesis (DAA) to 36 DAA. Type-II had the highest ratio of Gli-to-Glu from 20 to 36 DAA, and type-III ranked the second and without significant difference with type-I at 20 DAA. Glutenin contents were presented with the order of type-I > type-II > type-III from 28 to 36 DAA. The accumulation rates of glutenin and gliadin contents at the medium-late filling stage were different among cultivars, which resulted in the difference of raito of Gli-to-Glu. The HMW-GS content was higher in type-I than in type-II and type-III at 12 DAA, whereas showed the order of type-II > type-III > type-I from 20 DAA to maturity. The glutamine synthetase activity in flag leaf had positive correlations with glutenin content (P < 0.01) and ratio of HMW/LMW (P < 0.05) at 12 DAA and 20 DAA, but a negative correlation with the ratio of Gli-to-Glu at 20 DAA (P < 0.05). The endopeptidase activity in flag leaf had positive correlations with glutenin content (P < 0.01), ratio of HMW/LMW (P < 0.01), and gliadin content (P < 0.05) at 20 DAA and 28 DAA. High GS activity at the early-medium filling stage and high EP activity at the medium-late filling stage resulted in high contents of glutenin and gliadin, high ratio of HMW/LMW, and low ratio of Gli-to-Glu, which is favorable for high processing quality of the first-class strong-gluten wheat.

Estimation of Use Efficiency of Electrons in Fixation of CO₂ and Photorespiration for Maize (Zea mays) and Sorghum (Sorghum bicolor) under Photorespiratory Conditions

KANG Hua-Jing, TAO Ru-Liang, WANG Li-Xin, XIE Zi-Piao, LI Gong

Acta Agron Sin. 2011, 37(11):  2039-2045.  doi:10.3724/SP.J.1006.2011.02039

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Gas exchange and chlorophyll fluorescence for maize (Zea mays L.) and sorghum (Sorhumbicolor L.) at 380 μmol CO₂ mol^-1 and 30℃under photorespiratory condition, using a gas analyzer Li-6400, were measured. The results showed that the light response curve and rapid light curve (RLC) were well simulated by a modified rectangular hyperbola model. Calculation based on the simulated results indicated the electron flows to fix CO₂ for maize and sorghum were 198.60 and 178.00 μmol m^-2 s^-1, with the rate of 75.34% and 74.81%, respectively. The electron flows in photorespiration of maize and sorghum were 7.04 and 7.84 μmol m^-2 s^-1, with the rate of 2.67% and 3.29%, respectively. While by method of Valentini and Epron, the electron flows to fix CO₂ of maize and sorghum were 217.92 and 188.54 μmol m^-2 s^-1, with the rate of 82.68% and 79.24%, respectively, and those in photorespiration of maize and sorghum were 45.67 and 49.40 μmol m^-2 s^-1, with the rate of 17.32% and 20.76%, respectively. The results obtained by the former method showed that some electrons via PSII were used to CO₂ assimilation and photorespiration, whereas others associated with electron-consuming processes (e.g. O₂ acceptor cycle or water-water cycle) which should not be ignored, and the electron-consumption in this process was not constant under photorespiratory conditions. While the electron flows via PSII in photorespiration of maize and sorghum were overestimated by the latter method. The value of electron flows in photorespiration calculated by the latter method was about six times higher than those by the former. This is very important to evaluate the effect of photorespiration for plant protection.

Physiological Characteristics of Photosynthesis in Transgenic Wheat with Maize C₄-PEPC Gene under Field Conditions

TUN Qiong, HU Wei-Gang, LI Yan, JI Hua-Li, HU Lin, ZHANG Lei, HAN Lin-Lin

Acta Agron Sin. 2011, 37(11):  2046-2052.  doi:10.3724/SP.J.1006.2011.02046

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Transformation and high efficiency expression of key enzymes gene of C₄ photosynthetic characteristics are the important approach to improve photosynthetic efficiency and seed yield for C₃ crop wheat. Phosphoenolpyruvate carboxylase (PEPC) plays an important role in C₄ pathway. The authors have obtained several transgenic wheat lines with maize C₄ type PEPC gene. To explore the expression and functional characteristics of PEPC gene in transgenic wheat plants, PCR test, PEPC activity, gas exchange parameters, chlorophyll fluorescence parameters and yield related characteristics of transgenic wheat lines were determined, using untransformed wheat as the control, and the value of diurnal photosynthesis cumulative (VDPC) at the four stages and the correlations between net photosynthesis rate and stomata conductance, intercellular CO₂ concentration and transpiration rate were analyzed. The results showed that in the four determination stages, net photosynthetic rate of transgenic wheat obviously increased For instance VDPC was increased by 29.1% and 23.3% on the 15th day after anthesis. Stomatal conductance and transpiration rate in flag leaves of transgenic wheat obviously increased compared to the control, and intercellular CO₂ concentration decreased. Net photosynthetic rate of transgenic wheat enhanced by 23.2%, and variation scope of F_v/F_m, q_p, NPQ, and Φ_PSII of transgenic wheat was smaller than that of the control from 8:00 to 12:00 on the 15th day after anthesis; the major yield characteristics, such as weight per stem, thousand grain weight, single spike weight and harvest index of transgenic wheat, were significantly higher than those of the control. The results described above indicated that the transgenic wheat expressing maize PEPC gene showed better photosynthetic characteristics than untransformed plant and has the potential to increase wheat yield.

Photosynthetic Characteristics after Anthesis in Flag Leaves of Wheat Cultivars Xiaoyan 54 and 8602 and Their Hybrids Xiaoyan 41 and Xiaoyan 81

ZHANG Wen-Ting, SUN Xiao-Lin, BANG Qin, ZHANG Xi-Bin, YANG Xin-Hong, MENG Qiang-Wei, ZHAO Shi-Jie

Acta Agron Sin. 2011, 37(11):  2053-2058.  doi:10.3724/SP.J.1006.2011.02053

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Wheat lines Xiaoyan 41 (XY41) and Xiaoyan 81 (XY81) are hybrid progenies of Xiaoyan 54 (XY54) and 8602, which show distinct differences from their parents in photosynthesis characteristics. In this study, the gas exchange, chlorophyll fluorescence, and 820 nm transmission in flag leaves after anthesis were compared between hybrids XY41 and XY81 and parents XY54 and 8602. At anthesis, contents of chlorophyll a, chlorophyll b, and carotenoids were higher in the hybrids than in the parents, especially in XY81. Photosynthetic rate (P_n) and carboxylation efficiency (CE) were the highest in 8602, whereas stomatal conductance (Gs) was the lowest in XY54, followed by XY41. The four cultivars had similar values of intercellular CO₂ concentration (C_i), ratio of variable F_K to the amplitude F_J–F_O (W_k), maximum quantum yield of PSII (F_v/F_m), efficiency that an electron moves to Q_A downstream (Ψo), efficiency with which an electron from the intersystem electron carriers moves to electron acceptors in reduced end at the PSI acceptor side (δ_Ro), relative variable transmission at 820 nm (ΔI/I_o), actually photochemical efficiency of PSII (Φ_PSII), and nonphotochemical quenching (NPQ). After anthesis, the decreases of pigment contents, P_n, CE, G_s, F_v/F_m, δ_Ro, and ΔI/I_o and the increase of W_k were larger in XY54 than in other cultivars. No differences were detected in C_i, Ψ_o, Φ_PSII, and NPQ among cultivars, and these parameters varied similarly in 8602 and the hybrids during the whole developmental period tested. Compared with XY 54, XY41, and XY81 had slower decrease in P_n, and this resulted from the difference in dark reaction capacity rather than chlorophyll contents, stomatal conductance, and photosynthetic light reaction capacity. XY41 and XY81 might inherit the high dark reaction capacity from parent 8602.

T6VS?6AL Chromosome Translocation Does Not Alter Accumulations of High Molecular Weight Glutenin Subunits and Glutenin Macropolymer in Wheat Grain

XU Tian-Tian, CA Jian, HONG Bei, QI Ceng-Jun, DAI Ting-Bei, CAO Wei-Xing, JIANG Dong

Acta Agronomica Sinica. 2011, 37(11):  2059-2065.  doi:10.3724/SP.J.1006.2011.02059

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 Four lines from an F₈ recombinant inbred line (RIL) population were identified and classed into the the T6VS·6AL and non the T6VS·6AL groups. The grain protein content differed between the groups. The time-course changes in content and accumulation of HMW-GS and GMP during grain filling were detected. All HMW-glutenin subunits were detected at 13 d after anthesis (DAA) in grains of the four lines. However, the initial formation time differed within subunits. The content and accumulation of the HMW-GS increased during the whole grain filling period, and the rapid accumulation occurred from 23 DAA to maturity. The content and accumulation of HMW-GS and GMP showed no significant differences between the two groups. However, content and accumulation of HMW-GS of the high grain protein content lines were higher than those of the contrasting low protein content lines. In addition, grain gliadin content was found higher in the lines containing the T6VS·6AL chromosome translocation segment than in the contrasting lines.

Effects of Two Row Spaces and Intercropping on Forage and Crude Protein Yields of Oat (Avena sativa L.) and Common Vetch (Vicia sativa L.)

CHEN Gong, GUO Li-Mei, LIN Chang-Zhong, GUO Lai-Chun, DIAO Guo-Jun, HU Ti-Gao, CENG Zhao-Hai

Acta Agron Sin. 2011, 37(11):  2066-2074.  doi:10.3724/SP.J.1006.2011.02066

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Intercropping of oat (Avena sativa L.) and common vetch (Vicia sativa L.) is a widely used forage production system in Northwestern China. Due to different cropping managements, whether intercropped crops can yield higher than mono-cropped oat was not consistent in previous studies. This experiment was carried out at Baicheng Academy of Agricultural Science in Jilin province in 2009–2010. The quantity and quality of forage yield were justified with mono-cropped oat (B1) and common vetch (B2) as well as their intercropping (B3) under two different row spaces (A1: 33 cm and A2: 16.5 cm). In 2009, seeding rate of each row was the same for A1 and A2, so the total seeding rate for A2 was doubled; in 2010, the total seeding rate was the same for A1 and A2. The results showed that dry matter yields under intercropping system increased by 24% and 30% compared with those under the mono-cropping of oat and common vetch respectively. Crude protein yield in intercropped system was much higher (100%) than that in mono-cropped oat, but 20% less than that in mono-cropped vetch. A2 produced 13% and 29% more forage yield in 2009 and 2010 than A1. The effects of row spaces were less noticed in crude protein yield.With 16.5 cm of row spaces, oat seeding rate at 87.5 kg ha^–1 and common vetch seeding rate at 75 kg ha^–1, highest forage yield of 19.8 t ha^–1 and crude protein yield of 2.43 t ha^–1 were achieved with LER 1.55. Intercropping of oat and vetch could be a promising intercropping system in the studied area and the mentioned seeding rate and row space would be the best combination for forage production.

Photosynthetic Characteristics of the Isonuclear Alloplasmic CMS and Maintainer Lines of Dian-type I in Japonica Rice

SU Jia-Xiu, TAN Hua-Lin, XU Jin, LI Wei-Hua, HAI Mei-Rong, WANG Ting

Acta Agron Sin. 2011, 37(11):  2075-2084.  doi:10.3724/SP.J.1006.2011.02075

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CMS and maintainer line of Dian-type Iis one of the most important technological systems used for japonica hybrid rice in China. Using Li-6400 portable photosynthesis system, seven photosynthetic parameters, including net photosynthetic rate (P_n), stomatal conductance (G_s), intercellular CO₂ concentration (C_i), transpiration rate (T_r), water use efficiency (WUE), stomatal limitation (L_s), and chlorophyll content(SPAD), were measured on the top three leaves of 79 pairs of isonuclear alloplasmic CMS and maintainer lines of Dian-type I of Japonica rice. The results showed that there were significant differences of the parameters among the CMS lines and maintainers. The difference among genotypes is a valuable genetic basis for improvement of photosynthetic efficiency of CMS and maintainer lines. There were positive correlations significantly between CMS and maintainer lines on the seven parameters. Although there were no significant differences between CMS and maintainer lines on six photosynthetic parameters, but average value of SPAD of CMS lines was greater than that of maintainer lines significantly, which indicated that SPAD value was not a limiting factor for photosynthesis, and Dian-type I cytoplasm had no negative effect on photosynthesis, and chlorophyll content of CMS lines developed based on Dian-type I cytoplasm could be increased. According to the results of clustering analysis on flag leaf photosynthetic parameters, the CMS and maintainer lines could be classified into two types with high and lower photosynthetic rates, respectively. The results of this study could provide a theoretical basis for development of Dian-type I CMS lines with high efficiency of photosynthesis, and be used as theoretical references for development of japonica hybrid rice with high efficiency of photosynthesis.

Effects of Salt Stress on Photosystem II Activity in Sweet Sorghum Seedlings Grown in Pots Outdoors

WANG Cai-Juan, LI Zhi-Jiang, WANG Xiao-Lin, JIANG Chen-Dao, TANG Yu-Dan, GU Wei-Ban, DAN Lei

Acta Agron Sin. 2011, 37(11):  2085-2093.  doi:10.3724/SP.J.1006.2011.02085

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The effects of the different NaCl concentration treatments (100 mmol L^–1, 200 mmol L^–1) on photosynthetic characteristics and the photosystem II (PSII) activity in sweet sorghum seedlings grown in pots outdoors were carefully investigated in this study. The Na⁺ content and the ratio of Na⁺/K⁺ in leaves increased significantly with increases of NaCl concentration and treatment time. The net photosynthetic rate (P_n), the efficiency of excitation energy captured by open PSII reaction centers (F_v¢/F_m¢), the photochemical quenching (q_P) and the actual photosystem II efficiency (Φ_PSII) all decreased while the non-photochemical quenching (NPQ) increased greatly with the increasing of NaCl concentration. In addition, P_n, F_v¢/F_m¢, q_P and Φ_PSII had an ameliorative trend in the 100 mmol L^–1 NaCl treatment, but this phenomenon did not appear in 200 mmol L^–1 NaCl treatment. The maximum quantum yield of photosystem II photochemistry (F_v/F_m) was slightly affected in 100 mmol L^–1 NaCl treatment, whereas distinct decreased in 200 mmol L^–1 NaCl treatment. The chlorophyll fluorescence kinetic curves exhibited no changes at the beginning of NaCl treatment, while the rising speeds of O-K and O-J phases increased markedly in the 200 mmol L^–1 NaCl treatment after five days. Furthermore, the photoinactivation rate of PSII did not change in both treatments while the repair rate was depressed significantly after five days treatment of 200 mmol L^–1 NaCl. Therefore, we suggest that under salt stress the decrease of carbon assimilation capability in leaves of sweet sorghum seedlings grown in pots outdoors is mainly due to the accumulation of Na⁺, and salt stress alters the excited energy distribution. During salt stress outdoors, the decline of PSII activity is majorly attributed to the depression of the repair rate of the photoinactivated PSII, rather than the photoinactivation rate under high light. It is helpful to understand the mechanisms of photoinhibition in C4 crop outdoors under salt stress to some extent.

Differences of Potassium Efficiency Characteristics and Root Morphology between Two Cotton Genotypes

HAO Yan-Chu, JIANG Cun-Cang, WANG Xiao-Li, JIA Ying, CHEN Fang

Acta Agron Sin. 2011, 37(11):  2094-2098.  doi:10.3724/SP.J.1006.2011.02094

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The objective of this study was to analysis K-efficiency and root morphology in two kinds of K-efficiency cotton genotypes. The results showed that K-efficiency of genotype 103 was 160.2 g g^-1 and 47.5 g g^-1 at low (4 mg L^–1) and high (40 mg L^–1) K levels while genotype 122 was 133.8 g g^-1 and 37.9 g g^-1. In addition, genotype103 had higher ability to transfer K to leaves than genotype 122. Under two K levels, Leaves K accumulation of genotype 103 was 84.6% and 62.6% of the whole plant, while that of genotype 122 was 63.4% and 56.0%. High K-efficiency genotype 103 had higher total root length, surface area and volume than low K-efficiency genotype 122 regardless of the K concentration treated. Especially when the concentration of K in the environment was poor, genotype103 still had better root system than genotype122. When K was insufficient, the total root length and total root surface area of genotype 103 increased by 46.2% and 13.9%, while genotype 122 showed decreases in root parameters. Root system was classified into fine roots, middle roots and coarse roots according to root diameter. Low K condition limited the development of coarse roots. In genotype 103, coarse root length, surface area and root volume decreased by 54.0%, 62.8%, and 75.2% respectively, meanwhile, genotype 122 also had a reducing coarse root parameters. Effect of low K condition on fine roots varied in different cotton genotypes. The fine root length, surface area and volume of genotype 103 increased by 69.0%, 77.0%, and 80.4%, compared with these in appropriate K treatment, and were 1.9, 2.3, and 2.6 times higher than these of genotype 122, respectively.

RESEARCH NOTES

Effects of Plant Density on Ear Leaf Anatomical Structure in Summer Maize

LI Chun-Ai, WANG Ting-Liang, CHENG Xiang-Wen, CAO Re-Yao, LI Yun, HU Feng, LI Chao-Hai

Acta Agron Sin. 2011, 37(11):  2099-2105.  doi:10.3724/SP.J.1006.2011.02099

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A field experiment was conducted using maize hybrid Zhengdan 958 and Xundan 20 in 2006–2008, combined with paraffin method and ultramicrotomy to reveal relationship between plant density and ear leaf anatomic structure. The results showed that all the leaf thickness, vein area in cross section and xylem areain cross section, the stomata frequencies of upper and lower epidermis (for hybrid Zhengdan 958 the difference is significant, but for Xundan 20 is not),the number of grana in the chloroplast of mesophyll and chloroplasts in the vascular bundle sheath were decreased with the increase of plant density;the yield was the highest at the plant density of 75 000 plants ha^-1.The xylem area in cross section of leaf vein, stomata frequencies, number of grana in the chloroplast of mesophyll and chloroplasts in the vascular bundle sheath had remarkably negative correlation with plant density,but positive correlation with the kernel weight and grainnumber of single ear and 1000-kernel weight. The leaf structural parameters and agronomic indices will be important for application in maize production in the future.

Functional Analysis of TNBL1 Gene in Wheat Defense Response to Barley yellow dwarf virus Using BSMV-VIGS Technique

ZHAO Dan, ZHAO Ji-Rong, HUANG Qian, LI Ning, HUANG Tie-Jing, ZHANG Ceng-Yan

Acta Agron Sin. 2011, 37(11):  2106-2110.  doi:10.3724/SP.J.1006.2011.02106

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Barley yellow dwarf virus (BYDV), transmitted by at least 25 species of aphids, causes one of the most serious virus diseases of wheat worldwide. Through cDNA-AFLP analysis, we identified a cDNA fragment with 292bp expressing in the BYDV-resistant wheat-Thinopyrun intermedium translocation line YW642, but not in susceptible wheat Zhong8601. The full-length cDNA sequence of the gene, namely TNBL1, was cloned by RACE and RT-PCR methods, which encodes a putative NBS-LRR protein. This study focused on the functional analysis of TNBL1 in wheat defense to BYDV infection using Barley stripe mosaic virus (BSMV)-based virus-inducing gene silencing method. After the specific fragment of TNBL1 was addedwith 2 restriction-enzyme sequences byPCR, and digested and ligased with the digested BSMV-γ, the recombinant BSMV-γ:TNBL1as construct was obtained. The three components of the BSMV-VIGS vectors, BSMV-TNBL1as、BSMV-α and BSMV-β were transcribed in vitro, and mixed with equal quantity and inoculated onto the first and second leaves of the resistant line YW642 seedlings at the two-leaf stage. As a result, the TNBL1 expression was obviously repressed (silenced) in YW642 treated by BSMV:TNBL1. These seedlings were further inoculated with BYDV aphids. The BYDV content was much higher in the TNBL1-silenced YW642 plants than that in the control YW642 plants without BSMV:TNBL1 treatment. Furthermore, the TNBL1-silenced YW642 plants were susceptible to BYDV infection with the viral symptom. These results indicated that the TNBL1 gene is an important gene positively involved in wheat defense response to BYDV infection.

Relationship between Pericarp Structure and Kernel Tenderness in Super Sweet Corn

LE Su-Ju, XIAO De-Xin, LIU Feng-Fei, CENG Mu-Heng, WANG Wei-Quan, WANG Xiao-Meng

Acta Agron Sin. 2011, 37(11):  2111-2116.  doi:10.3724/SP.J.1006.2011.02111

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In order to understand the relationship between kernel tenderness and pericarp structure, histological studies of a set of super-sweet corn (Z. mays subsp. mays) inbred lines (T4, T5, T19, T38, and T39) kernels at different developmental stages were conducted. The result indicated thatsuper-sweet cornpericarp was formed with several layers of cells, where the outermost layer of cells packed tightly with horny cell walls. T38 and T39 inbred lines had a good eating qualities due to their thinner cell walls and less cell layers of pericarp combining with lower level of fibrosis and lignification of cell wall. On the contrary, T4 and T5 inbred lines did not have a eating qualities with thicker cell walls and more cell layers of pericarp combining with high level of fibrosis and lignification of cell wall. T19 had the worst taste for its high-density pericarp cells adding very high fibrosis and lignification of cell wall among the five inbred lines.

Detection of Genetically Modified Maize MON88017 by Quantitative Real-Time PCR

YUAN Lei, SUN Gong-Wei, LI Fan, LI Ning, ZHAO Lei, LU Xin-Bei

Acta Agron Sin. 2011, 37(11):  2117-2121.  doi:10.3724/SP.J.1006.2011.02117

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Transgenic event-specific primers and Taqman probes based on the left-flanking sequence of MON88017 and endogenous gene (zSSIIb) were designed, and a real-time PCR method was developed to quantitatively detect the event-specific genetically modified maize. The reference molecule composed of endogenous gene and flanking sequence of MON88017 was constructed artificially. Two standard curves of the reference gene sequence and the 5' flanking sequence were established. We detected five mixed samples their genetically modified contents of MON88017 were 0.01%, 0.05%, 0.1%, 0.5%, and 1% respectively. The lowest detection limit was 19–30 copies. This study indicated that the Taqman probes real-time PCR is highly specific and sensitive.