Frontiers of Agriculture in China

ISSN 1673-7334

ISSN 1673-744X(Online)

CN 11-5729/S

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Research articles
Mapped SSR markers unevenly distributed on the cotton chromosomes
Zhongxu LIN, Daojun YUAN, Xianlong ZHANG,
Front. Agric. China. 2010, 4 (3): 257-264.

Abstract   PDF (269KB)
In the past 15 years, more than 9000 SSR markers have been developed, which are a valuable resource for cotton genetics and breeding. Many of them have been mapped in cotton, and several high-density genetic maps have been constructed. In this paper, we jointly analyzed the mapped SSRs to characterize their motifs on cotton chromosomes. The results showed that SSR loci were distributed unevenly on cotton chromosomes with more on C11 and C19 and less on C02 and C04. Different sources of SSRs contributed different predominant SSR types, which were also distributed unequally on cotton chromosomes. Compared to SSRs derived from enriched libraries and BAC end sequences, EST-SSRs could generate novel SSR motifs especially for AT/TA of di-, tetra-, penta- and hexanucleotide. SSR motifs showed a biased distribution on cotton chromosomes with more on C05, C11, and C19, and less on C02 and C04. The uneven distribution of SSRs on cotton chromosomes would help us to enrich the cotton genetic map by motif-specific SSRs. This analysis gave us new knowledge on the architecture of the cotton genome.
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Genetic analysis of tolerance to aluminum toxin at seedling stage in soybean based on major gene plus polygene mixed inheritance model
Paul C. KORIR, Jinshe WANG, Tuanjie ZHAO, Junyi GAI,
Front. Agric. China. 2010, 4 (3): 265-271.

Abstract   PDF (166KB)
The segregation analysis based on major gene plus polygene mixed inheritance model was employed to study the inheritance of aluminum (Al) tolerance in soybean. A recombinant inbred line (RIL) population derived from a cross between Al-tolerant (KF No.1) and Al-sensitive (NN 1138-2) parents along with both parents was analyzed using phenotypic data of three traits, i.e. relative total plant dry weight (RTDW), relative shoot dry weight (RSDW) and relative root dry weight (RRDW) assayed in sand culture. Significant difference among RILs was observed, and frequency distributions showed characteristics of mixed distributions, which suggested that inheritance of Al tolerance conformed to major gene plus polygene mixed inheritance model. The results indicated that the genetic model type I (four additive major genes plus additive polygenes) was the most fitting one for the traits, indicating some common genetic mechanism lying among the traits, though their respective best fitting models were somewhat different, i.e. I-6, I-9 and I-8 for RTDW, RSDW and RRDW, respectively, within model type I. It supports to choose RTDW as the representative for Al tolerance evaluation in addition to its virtue of being the whole-plant trait and easy to measure in sand culture. Averagely, the major genes contributed about 50%, while the collective polygenes contributed about 30% to the phenotypic variation, indicating both being important in breeding for Al tolerance. The study recommends that the segregation analysis can be used by soybean breeders who have accumulated abundant segregation data to obtain preliminary information on the inheritance of their breeding materials, and can be used as a first screening for presence of major genes followed with molecular marker analysis to further confirm and locate the major genes of quantitative traits.
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Identification of phosphorus starvation tolerant soybean ( Glycine max ) germplasms
Xihuan LI, Wensuo CHANG, Caiying ZHANG, Junyi GAI,
Front. Agric. China. 2010, 4 (3): 272-279.

Abstract   PDF (324KB)
In the present study, 156 soybean genotypes mainly from the Hebei growing-area were identified for their low phosphorus starvation tolerance. The results showed that the relative values (value of dry wt. in zero soil P/value of dry wt. in adequate soil P) of shoot dry weight and root dry weight of all soybean varieties, at the phosphorus concentration of 0 mmol·L−1 and 1.0 mmol·L−1 in supplied solution, offered a quick and reliable method to identify and classify genotypes for phosphorus starvation tolerance. We describe three classes: tolerant, moderate and sensitive P starvation groups which had 29, 59, and 68 cultivars, respectively. Eight varieties, Ji-dou11, Lü75, Hei-da-li, Zhong-huang15, Zhe98-14, Da-mao-jiao, Da-huang-dou and Zha-lai-te-qi were screened out for their high tolerance to phosphorus deficiency under zero soil phosphorus conditions. In addition, the relationships between soybean phosphorus starvation tolerance and seed phosphorus content, root morphology, rhizosphere acidification are also discussed.
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A study on indicators and evaluation stages of aluminum tolerance in soybean
Paul C. KORIR, Tuanjie ZHAO, Junyi GAI,
Front. Agric. China. 2010, 4 (3): 280-286.

Abstract   PDF (138KB)
To determine an appropriate indicator and a suitable stage for evaluating tolerance of soybeans to aluminum (Al) toxin is one of the keys to effective breeding for the trait. Seventeen accessions selected as tolerant from a previous test program by using average membership index (FAi) as indicator, plus one tolerant (PI.416937) and one sensitive (NN1138-2) check, were assayed in sand culture pot experiments, totaling four experiments, each for evaluation at V3, V5, V7 and V9 stage, respectively, each in a randomized complete block design with three replications, and each genotype exposed to two Al levels (0 and 480 µM). The relative values of shoot dry weight (RSDW), root dry weight (RRDW), total plant dry weight (RTDW), total root length (RTRL) and total root surface area (RRSA) as the tolerance indicators as well as FAi were compared. All the indicators showed significant variation in Al tolerance among genotypes over and across the leaf stages, but Genotype × Stage interactions were significant only for RTRL and RRSA, indicating that they were less stable among stages than RTDW, RSDW and RRDW. Among the latter three, RTDW was chosen as the major indicator of Al tolerance due to its relatively better stability, higher correlation with other indicators and easier measuring procedure than the others. The seedling age applicable for screening was not definitive, but V5 appeared to compromise between time spent resulting from screening the relatively older seedlings at later stages and low variation among genotypes at a younger stage. The differences of Al tolerance among the tested accessions were further detected by using RTDW, and superior Al tolerant accessions identified were PI.509080 (South Korea), N23533 and N24282 (Northeast China) and PI.159322 (USA), comparable to the putative tolerant check PI.416937 (Japan) at all vegetative stages.
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Advances in the studies of Rice stripe virus
Donglai XIAO, Weiming LI, Taiyun WEI, Zujian WU, Lianhui XIE
Front. Agric. China. 2010, 4 (3): 287-292.

Abstract   PDF (119KB)
Rice stripe virus (RSV), the type member of the genus Tenuivirus, is one of the most economically important pathogens of rice and is repeatedly epidemic in China, Japan and Korea. The latest achievements of the studies on the biological functions of virus-encoded proteins, pathogenicity differentiation and genetic diversity of virus, virus-plant host interactions and management of virus were reviewed. The current problems encountered during studies and some approaches for further research were discussed.
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One magic pteridophyte ( Pteris vittata L.): Application in remediating arsenic contaminated soils and mechanism of arsenic hyperaccumulation
Jiemin ZHENG, Tianxin NIU, Genliang WU, Wenyue CHEN,
Front. Agric. China. 2010, 4 (3): 293-298.

Abstract   PDF (112KB)
One pteridophyte, Pteris vittata L., was found to be a suitable plant used in phytoremediation of the soils contaminated with excess arsenic. In this paper, literature on this fern’s potential in remediating soils contaminated with arsenic, its possible detoxification strategies, and the measures facilitating its practical application in phytoremediation were reviewed. Some of the unresolved questions about this fern in both detoxification and phytoextraction of arsenic (As) were also listed. The aim of this paper was to introduce this magic fern to more researchers focusing on a wider field of discipline and make it more useful in both macroscopic and microcosmic science world.
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Heterosis and combining ability analysis for yield and its components in Indian mustard ( Brassica juncea L. Czern & Coss)
Priti Gupta, H B Chaudhary, Sandeep Kumar Lal,
Front. Agric. China. 2010, 4 (3): 299-307.

Abstract   PDF (126KB)
Half diallel analysis of eight parents was carried out to identify the high heterotic crosses and their relationship in terms of general and specific combining ability (GCA & SCA) in Brassica juncea L. Czern and Coss at IARI, New Delhi, during 2007–2008 and 2008–2009. The relative heterosis and heterobeltiosis were observed to be the highest with respect to seed yield per 100 siliquae and days from sowing to 50% flowering in case of cross IC-199715 × IC-199714, EC-289602 × Prakash in the number of primary branches per plant and harvest index, Agra Local × Pusa Bahar in the length of main axis, Poorbijaya × Agra Local in the number of siliquae on main axis and EC-289602 × Pusa Bahar in the biologic yield per plant and seed yield per plant. Different cross combinations exhibited the maximum value of better and mid-parent heterosis for the remaining traits, viz., days to maturity, number of secondary branches per plant, plant height and 1000-seed weight. GCA and SCA variances were significant in all characters. The variance of GCA (s2g) was observed to be higher from sowing to 50% flowering and maturity in plant height and 1000-seed weight, whereas the variance of SCA (s2s) was higher in seed yield and other remaining parameters.
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Identification of senescence-related genes by cDNA-AFLP in cotton ( Gossypium hirsutum L.)
Chengjin GUO, Jinfeng ZHAO, Cundong LI, Kai XIAO, Haina ZHANG, Chuanfan SUN, Juntao GU,
Front. Agric. China. 2010, 4 (3): 308-316.

Abstract   PDF (161KB)
Premature senescence at the late developmental stage occurs frequently in cotton (Gossypium hirsutum L.) production in North China. It is desirable to develop elite cotton cultivars with non-premature senescence and high photosynthetic capacity. In this study, cDNA-amplified fragment length polymorphism (cDNA-AFLP) analysis was employed to identify the genes that are related to senescence in cotton. Using 64 primer combinations, about 3000 cDNA fragments were generated, and among them 42 had a markedly up-regulated expression pattern with the leaf growth progression. Based on cloning, sequencing, and Blast search analysis, it was determined that 24 TDFs with putative known biologic functions could be classified into several major categories, such as signal transduction, transcription regulation, stress-responsive, primary and secondary metabolism, nutrients recycling, photosynthesis, cell wall biosynthesis, and senescence-related. TDF31, TDF32 and TDF33, with high similarity to the senescent-regulating genes MAP kinase 9 (MKK9) and non-yellowing protein 1(NYE1) from Arabidopsis and bean senescence-associated receptor-like kinase (SARK) could play possible roles in responding or modulating the leaf senescence in cotton. Therefore, leaf senescence in cotton is a complicated network involving many biological processes. Some putative genes with important modulation functions in regulating or responding to the senescence need to be further analyzed.
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Cloning and prokaryotic expression in TaCaM2-3 of wheat and preparation of antiserum
Wei LIU, Aihua YAN, Chunyan HOU, Dongmei WANG,
Front. Agric. China. 2010, 4 (3): 317-322.

Abstract   PDF (184KB)
Multiple calmodulin (CaM) isoforms exist in plant organisms and vary by their primary structures of 148 amino acids. They have different expression patterns and/or target enzyme activation abilities. To further understand the biological significance of TaCaM isoforms, total RNA was isolated from mature leaves of wheat and then TaCaM2-3 gene was amplified by PCR after reverse transcription. The PCR product was generated into T-easy vector to subsequently sequence. Then the recombinant expression vector (pET28a-TaCaM2-3) was constructed and transformed into E. coli strain BL21 to obtain a high level expression vector of CaM. SDS-PAGE analysis showed that the recombinant E. coli could express an approximate 20 kD protein. A western blotting analysis showed an anti-CaM monoclonal antibody specifically bound to the 20 kD band of expressed product. TaCaM-II was purified by Ni-NTA affinity chromatography from recombinant bacterial lysate. TaCaM-II protein was used to immunize New Zealand white rabbits to produce a polyclonal antiserum. The specificity of the anti-TaCaM-II antiserum was successfully verified by western blotting analysis.
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The effects of phlorizin stress on the protective enzyme and metabolic regulation substances in the root of M. micromalus
Xiaojing CUI, Yanan WANG, Wenchao ZHEN,
Front. Agric. China. 2010, 4 (3): 323-327.

Abstract   PDF (88KB)
The effects of phlorizin stress on the protective enzyme (PE) and metabolic regulation substances (MRS) in the Malus micromalus root were studied by tissue culture method. Under four concentration gradients of phlorizin stress, the activity of superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), the content of malondialdehyde (MDA), proline (Pro) and soluble proteins were tested. The results showed that phlorizin stress could lead to different changing tendencies for different detection indicators. The changes of SOD activity, POD activity and Pro content appeared more evident. The difference between the treatments under phlorizin stress and the control without phlorizin stress was not significant in the initial period. With the extension of processing time the difference became more and more evident and the protective enzyme activities and metabolic regulation substances were influenced greatly under different concentrations of phlorizin stress. When the concentration of phlorizin got to 1g·L−1, the effect was the most obvious. The activities of protective enzymes were significantly improved, with the MDA production reduced, and Pro and soluble protein (SP) accumulation accelerated.
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Construction of an engineering strain expressing cry7Ab7 gene cloned from Bacillus thuringiensis
Liguang WANG, Tinghui LIU, Wei GUO, Jianxin TAN, Weiming SUN, Yongxiang SUN,
Front. Agric. China. 2010, 4 (3): 328-333.

Abstract   PDF (190KB)
The genotype of Bacillus thuringiensis (Bt)strain GW6 isolated in China was identified, and a novel cry7Ab gene was found based on the results of PCRs using 40 pairs of cry universal primers. The cry7Ab gene was cloned by PCR and named as cry7Ab7 by the Bt Delta Endotoxin Nomenclature Committee (GenBank Accession No. FJ940776). The construction of a novel three-dimensional structure of Cry7Ab7 protein via homology modeling methods indicated that the Cry7Ab7 protein was different from other Cry7Ab proteins within the domain structures. The cry7Ab7 gene was inserted into the BamHI/SalI sites of E. coli expression vector pET21b and the Bt-E. coli shuttle vector pSXY422b to construct the recombinant plasmids pET21b-7Ab7 and pSXY422b-7Ab7, which were then transformed into E. coli BL21 and Bt HD73cry in order to obtain the E. coli transformant EC7Ab7 and the engineering strain Bt HD7AB, respectively. The bioassay results showed that Cry7Ab7 protein inclusion bodies in EC7Ab7 and the crystal proteins in HD7AB were highly toxic to the second-instar larvae of Henosepilachna vigintioctomaculata with the LC50 values of 1.167 μg·μL−1 and 0.779 μg·μL−1, respectively. Our results clearly indicated that cry7Ab7 gene can offer important benefits to research on Coccinellidae insect pest control.
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Thiolysis-HPLC analysis of proanthocyanidins in health foods and their materials
Ye LI, Huiling LI, Tong CUI,
Front. Agric. China. 2010, 4 (3): 334-340.

Abstract   PDF (222KB)
Proantocyanidins (PCs) found in plant products, are reported to be beneficial to human health. A thiolysis-HPLC method was developed to determine PC in health foods and the basic PC material, grape seed extract (GSE). PCs were quantitatively resolved by acid-catalysis in the presence of benzyl mercaptan as nucleophiles. The reaction solutions of actual purchased products were injected into an HPLC and the gallic acid (GA), catechin (CT), epicatechin (EC), epicatechin gallate (ECG), epicatechin benzyl sulfide (EC-S), and epicatechin gallate benzyl sulfide (ECG-S) were separated and detected. The HPLC was performed on a Hypersil BDS C18 (4.6 mm id × 250 mm, 5 μm) column at 35°C, formic acid (0.05%) was employed as mobile phase A, and methanol ∶ acetonitrile= 1∶2 (v/v) was employed as mobile phase B. The flow rate of gradient elution was 0.8 mL·min−1, and the DAD detection was carried out at 280 nm. The relative standard deviation of CA, CT, EC, ECG, EGS and ECG-S followed by repeated reactions and injections were between 0.4%–4.2%, and the PC levels in six health foods were between 6.9–161.3 mg·g−1 with the mean degree of polymerization (mDP) 2.2–4.2. In a GSE product, the PC content was 531.4 mg·g−1, and the mDP was 9.0, which although higher than other products tested, was far below the labeled, claimed, concentration. Data showed this method to be accurate and sensitive and can be used for the quality evaluation of actual health products containing PC components.
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Impact of different combined doses of fertilizers with plant growth regulators on growth, yield attributes and yield of mustard ( Brassica campestris cv. B 9 ) under old alluvial soil of Burdwan, West Bengal, India
Arnab BANERJEE, Jayanta Kumar DATTA, Naba Kumar MONDAL,
Front. Agric. China. 2010, 4 (3): 341-351.

Abstract   PDF (158KB)
Field experiments were conducted with mustard (Brassica campestris cv. B9) consecutively for three years (2005, 2006 and 2007) in the Crop Research and Seed Multiplication Farm of Burdwan University, Burdwan, West Bengal, India. In the first year, varietal screening of mustard under recommended dose of chemical fertilizer (100∶50∶50) were performed with seven mustard varieties during November 2005 to February 2006. In the second year, two experiments were conducted to study the effects of six different combined doses of chemical fertilizer and biofertilizer and six different levels of growth retardant cycocel. In the third year, six different levels of compost along with the best screened dose of growth retardant (CCC-300 ppm) and the best combined dose of biofertilizer and chemical fertilizer (3/4thchemical fertilizer:1/4th biofertilizer) from the previous year were applied to study the effects on agronomic traits and growth attributes of mustard, and the field data were analyzed statistically. The variety B9 recorded a significant higher seed yield along with other yield contributing factors, which was found to be superior to other varieties under old alluvial soil of Burdwan, West Bengal, India, in 2005. In 2006 and 2007, seed yield was found to be the best for CCC-300 ppm treatment and the best combined dose of biofertilizer and chemical fertilizer was found to be 3/4th chemical fertilizer:1/4th biofertilizer. In 2007, the best yield was given by the treatment of 7.5 t·hm−2 compost along with the best dose of growth retardant cycocel and the best combined dose of biofertilizer and chemical fertilizer.
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The dynamics and correlation between nitrogen, phosphorus, potassium and calcium in a hazelnut fruit during its development
Lihong WEI, Qiuxi ZHAI,
Front. Agric. China. 2010, 4 (3): 352-357.

Abstract   PDF (201KB)
The dynamics and correlation between nitrogen, phosphorus, potassium and calcium were studied during the fruit development of three crossbreeds of hazelnut. The results showed that the young fruit rapid growth stage and kernel development stage were the critical periods for the absorption and accumulation of these mineral elements. With three crossbreeds, the content of nitrogen followed such a pattern that decreased in the initial stage, remained constantly in the middle stage and increased markedly in the late stage; phosphorus followed a slightly different pattern where it decreased in the initial stage, also remained constant in the middle stage, and first increased rapidly and then decreased rapidly in the late stage; potassium followed the pattern where it decreased mildly in the initial stage, increased gradually in the middle and late stages; calcium followed the pattern where it decreased sharply in the initial stage, remained constant in the middle and late stages. Nitrogen, phosphorus and potassium in fruits showed a significant or very significant positive correlation in the development course of hazelnut fruit and there was a dynamic equilibrium of coordination among the three elements.
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In vitro micro propagation of “sand pear” Pyrus pyrifolia (Burm. f.) Nakai
Zafrul HAQ, Z. A. KALOO,
Front. Agric. China. 2010, 4 (3): 358-361.

Abstract   PDF (217KB)
This study was undertaken to standardize protocols for in vitro explant establishment, shoot multiplication and in vitro rooting for Pyrus pyrifolia, growing in the Kashmir valley. The explant used was nodal cuttings of 2–3cm size with one or two axillary buds. The highest explant establishment (over 90%) was observed during spring while the lowest (60%) explant establishment frequency was observed during winter season. The shoots sprouted from the axillary buds were excised and subcultured on MS basal medium. The highest shoot multiplication rate (130 shoots from a single nodal cutting) was obtained on MS medium supplemented with BAP 2.2 mg·L−1 and kinetin 1 mg·L−1 after four weeks. The shoots were inoculated for rooting in root induction medium of half strength MS salts supplemented with plant growth regulators like IBA, IAA, NAA, and 2, 4-D. However, the best rooting response (8.50 roots per shoot) was obtained in combination of hormones (IBA+ NAA) 0.25 mg·L−1 each. The rooted micro shoots were acclimatized under decreasing humidity regime (95% RH to 75% RH over a period of 5±1 weeks). The highest ex vitro plantlet survival percentage (56.50%) of Pyrus pyrifolia was observed on sand+ soil (1∶1 v/v).
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The role of leptin in bovine adipocyte fat metabolism in vitro
Cai ZHANG, Zijun YANG, Shuling NIU, Zhe WANG,
Front. Agric. China. 2010, 4 (3): 362-365.

Abstract   PDF (164KB)
To identify the effect of leptin on adipocyte, preadipocyte was isolated from male dairy calves and cultured in a monolayer; 5 μg·L−1 bovine leptin were added in culture liquid when the cell differentiated into adipocytes. The result showed that leptin decreased the deposition of triglyceride (TG) and increased the levels of hormone sensitive lipase (HSL) mRNA in cells. At the same time, leptin suppressed the levels of endogenous leptin mRNA, and the levels of leptin receptor (Ob-R) mRNA were raised significantly. It indicated that leptin may induce more fat metabolism directly.
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Genetic diversity analysis of black locust ( Robinia pseudoacacia L.) distributed in China based on allozyme markers approach
Juntao GU, Minsheng YANG, Jinmao WANG, Jun ZHANG, Haiyong LIANG, Li JIA,
Front. Agric. China. 2010, 4 (3): 366-374.

Abstract   PDF (238KB)
Using starch gel and polyacrylamide gel electrophoresis, the genetic diversity of 19 populations of black locust (Robinia pseudoacacia L.) distributed in China has been evaluated based on seven allozyme systems. Among the 14 enzyme loci studied, 12 of them showed polymorphic, giving a total of 40 alleles. The average allelic number (A) and the average effective allelic number (Ae) were 3.004 and 2.178, whereas the average of expected heterozygosity (He) and the observed heterozygosity (Ho) were 0.525 and 0.440, respectively. The total genetic differentiation (HT) in the 19 populations tested was 0.536, including 0.515 within population (HS) and 0.021 among populations (DST). The mean coefficient of genetic differentiation (GST) was 0.038, indicating that 3.8% of genetic differentiation was resulted from the populations, and the rest from those within the population. The genetic distance among the 19 populations was small, ranging from 0.015 to 0.065. Cluster analysis based on AVERAGE showed that populations with closer geographical distribution had higher genetic similarity than that in populations with far away distribution. There was no evident geographic differentiation pattern in Chinese black locust populations at a biochemical level.
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Arbuscular mycorrhizal fungi and dark septate endophyte colonization in bamboo from Northeast India
Panna DAS, Highland KAYANG,
Front. Agric. China. 2010, 4 (3): 375-382.

Abstract   PDF (478KB)
To address paucity of the mycorrhizal studies in bamboo, we have investigated arbuscular mycorrhizal fungal (AMF) distribution and dark septate endophyte (DSE) colonization on four species of bamboo from Northeast India. Bambusa tulda exhibits Arum type of AMF morphology, and other bamboo species have Paris type. AMF colonization was significantly higher than DSE colonization (P<0.05). Vesicular and DSE colonization exhibit a significant positive correlation with organic carbon and available phosphorus, respectively (P<0.05). Of 17 species isolated from Acaulospora, Ambispora and Glomus, 12 were isolated from Phyllostachys manii. Acaulospora tuberculata, A. rehmii, Glomus intraradices and G. tortuosum were the most frequently distributed species. Shannon diversity index was the highest in P. manii. Principal component analysis (PCA) plots and cluster analysis suggest that P. manii was the most dissimilar bamboo species in terms of mycorrhizal colonization and soil properties.
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18 articles