Seed protein content, controlled by quantitative genes, is a very important trait for soybean. To identify the soybean quantitative trait loci (QTL) of seed protein content, which provides a base for MAS breeding and elite genes-discovering, a cross was made between the elite cultivar Zhonghuang 13, which exhibits a comprehensive excellent performance and has the higher protein content and the variety Dongshan 69 which has the lower protein content. The BC2F2 and BC2F3 backcross populations were further developed to identify protein composite interval mapping (ICIM), two QTLs flanked by the marker interval Satt396~Satt180 and Sct_001~
Satt654 were both detected in the BC2F2 and BC2F3 families. In the BC2F2 families, 3 QTLs were found on A1, C1 and J linkage groups, which accounted for 10.00%, 7.07% and 9.23% phenotypic variance, respectively. In the BC2F3 families, 4 QTLs were identified on B1, C1, I and J linkage groups, which explained 17.57%, 3.46%, 8.53% and 11.80% phenotypic variance, respectively. Sct_001~Satt654 may be a new interval associated with protein content QTL.
This experiment was to construct the expression vectors which contain hrpZpsta and RIPs genes and transform these resistant genes into soybean genome via Agrobacterium-tumefaciens mediated transformation method. Soybean cultivar Jinong 28 cotyledonary node was used as explant. The putative transgenic plants were screened with salt-tolerance gene BADH and 18 T1 positive transgenic progenies were obtained. The PCR, Southern blot, RT-PCR analysis showed that hrpZpsta and RIPs genes had been transformed into soybean genome and could be inherited in the next progeny.
Drought stress seriously affects plant growth and development as well as reduces quality and productivity of crops. In order to gain some key functional genes in response to dehydration/drought stress, a digital gene Expression Tag profile of two drought tolerance and sensitivity materials under dehydration had been constructed in our previous study. In the research, a gene encoding bHLH transcription factor, namely GmbHLH25, was identified from expression profile. This gene was significantly induced in leaves and roots under dehydration and the expression showed an obvious difference between two materials. Expression level of GmbHLH25 gene was validated by real-time quantitative PCR. The CDS sequence of GmbHLH had been cloned from two materials. GmbHLH25 gene encodes 368 aa and contains one basic Helix-Loop-Helix(bHLH) motif comprised of 50 amino acid. The homology tree demonstrated that GmbHLH25 was at the same evolutionary branch with homologous proteins of Lotus corniculatus and Medicago. Subcellular localization revealed that GmbHLH25 was distributed preferentially to nucleus. PCR and RT-PCR results showed that GmbHLH25 gene had been integrated into tobacco genome. Over expression of GmbHLH25 gene improved drought and salt tolerance capacities under water-deficit and high salt conditions. This study will help to clarify the molecular mechanisms of plants response to drought.
Late embryogenesis abundant (LEA) proteins, a class of extremely hydrophilic proteins which were first identified in plant embryogenesis, were also found in vegetative tissues. They play important roles in plant desiccation tolerance. In this study, a genome-wide search was carried out to identify LEA genes in castor bean (Ricinus communis L.), a Euphorbiaceae plant that is well known for its plasticity to unfavorable environments. As a result, a total of 27 RcLEAs representing 8 groups were identified, 3 of which were found to have alternative splicing isoforms. Based on protein characteristics and phylogenesis, the 27 RcLEAs contained 0 to 2 introns and distributed across 24 released scaffolds, and were named as RcLEA1-1 and -2, RcLEA2-1 and -2, RcLEA3-1 to -3, RcLEA4-1 to -7, RcLEA5-1 and -2, RcLEA6-1 and -2, RcLEA7-1 to -5 and RcLEA8-1 to 4. Using the BLAT method, all RcLEA genes were expressed in at least one of the examined tissues such as leaf, flower, endosperm_II_III, endosperm_V_VI and seed. Their promoters enriched stress-responsive cis-acting elements including LTRE, ABRE, MYC, MYB and W-box.
Photosynthetic rate, chlorophyll content and enzyme activity of silique wall of rapeseed were investigated to study the mechanism on declining of rapeseed silique photosynthesis and its effect on yield. Materials included 3 cultivars as Zhongshuang 9 (ZS9), Zhongyouza 11 (ZYZ11)and Huayouza 14 (HYZ14). Results showed that net photosynthetic rate (Pn) decreased during silique growth. Similar responses were found in ribulose 1,5 - bisphosphate carboxylase (RuBPcase) content, soluble protein content, chlorophyll content, superoxide dismutase (SOD) and catalase (CAT) activities in silique wall. The progressive increase of malondialdehyde (MDA) occurred during the silique stage. Results showed that silique photosynthetic rate had significant correlations in second-degree parabola with contents of chlorophyll, soluble protein, MDA, and activities of SOD and CAT. Furthermore, RuBPCase had highly significant linear relationships with contents of chlorophyll, soluble protein, MDA, and activities of SOD。
To understand the nutrient uptaking during and after the waterlogging stress, a pot experiment was conducted to investigate the effects of different waterlogging durations on rapeseed growth and recovery using Brassica napus L. cv Zhongshuang 11. Results showed that the adverse effect of waterlogging on oilseed rape seedling was not only on growth during stress, but also on recovery period after flooding. The leaf nitrogen (N), phosphorus (P), potassium (K), iron (Fe), zinc (Zn) and copper (Cu) concentrations were significantly decreased, but the leaf manganese (Mn) concentration was significantly increased with the period of waterlogging lasted. The root N, Fe, Zn, and Cu concentrations were significantly increased, the root P concentration was significantly decreased, the root Mn concentration was significantly increased at 3d then significantly decreased, and the root K concentration was no significantly different during waterlogging. Moreover, in soils, the proportion of nitrate to ammonium and available P concentration were both significantly decreased. Available K, Fe and Zn concentrations were curved. Cu concentration in soils was no significantly different during waterlogging but significantly decreased after.
To understand the extra-large seed characteristics of Brassica napus var DL01, silique traits were investigated in field on growth period, using normal seed line C001 as control. Results showed that DL01 average growth period was 207d, which was ～17d longer than C001. Grain filling period was 12d longer. Stalk diameter as well as ovary length, width and weight were 1.30, 1.21, 1.39 and 1.46 folds of C001 respectively at the first day of flowering. 100 seed weight (1.20g) and volume, as well as filling rate were 2 fold. The maximum photosynthesis areas of silique pericarp and fresh weight of siliques were 2.2 and 3.0 fold respectively. But their embryo number per silique were similar, and seeds per silique could be 64% less than C001. Moisture content (MC), the peel MC was 3-5% higher, and the period of high MC (>80%) lasted 80 days, while the period of high MC (>75%) in C001 lasted only 68 days. The seed fresh weight of DL01 and C001 reached the peak at the 62nd and 47th day, with 50% and 52% MC respectively. In conclusion, the extra size of DL01 resulted from large ovary, thick stalk, and wide silique surface, as well as fewer grain number, higher MC of seed and silique, and longer filling duration.
In this study, the grafting method was used to create composite plants material with the same scion but different cotyledons and the parts below cotyledons. The cut-in grafting method was used to study the changes of cotyledon area from cotyledon stage (VC) to second-node (V2) stage. The widely-planting soybean varieties over the years in Yellow-Huai-Hai Rivers Valley of China were used as stock genotypes, and Zhonghuang 39 was as scion. 3 soybean varieties with different 100-seed weight were selected as test materials, and the post-emergence changes of cotyledons area and physiological indices and their effects on seedling growth and development in soybean by setting the shading and Normal sunshine treatments were studied. The results showed that cotyledon area increased during the seedling period from VC to V2 among all the tested varieties, with the increment range from 65.61% to 209.29%. The increment of cotyledon area from V2 to VC was negatively correlated with the cotyledon area at VC stage. The cotyledon area increment from VC to V2 decreased with the releasing years of the varieties. Beginning from Day 9 after emergence, the cotyledon area constantly declined in all varieties, and the contents of chlorophyll a, total chlorophylls and the dry weight of cotyledons decreased along with the seedling growth, the chlorophyll b content increased first and then decreased. Cotyledon soluble sugar content decreased for some days and then increased gradually, and finally decreased with aging cotyledons. After the shading, the changes of cotyledon area, dry weight, chlorophyll a, b and soluble sugar contents became more prominent than those in Normal sunshine treatment, indicating that limiting the cotyledon "source" functions could affect the synthesis, degradation and transport of chlorophylls and soluble sugars in cotyledons, and the changes of cotyledon area and their dry weight. Removing cotyledons had impacts on seedling growth and development, with shorter seedling height, slower development process, and lower seedling dry weight. It was concluded that cotyledon is one of the important vegetative organs of soybean at seedling stage. Cotyledons should be protected and effectively utilized in soybean management.
Variation of photosynthetic characteristics, leaf chlorophyll fluorescence characteristics and some physiological characteristics of four drought-resistant soybean varieties and four ordinary soybean varieties were investigated under drought stress. The results showed that the net photosynthetic rate（Pn）, stomatal conductance（Gs）and transpiration rate（Tr） of three growth stages were declined under drought stress. The ordinary soybean varieties were more declined than drought-resistant varieties. The intercellular CO2 concentration （Ci）, especially of the ordinary soybean varieties decreased significatly at R2 stage. The apparent mesophyll conductance（AMC） had no obvious change at R2. The Ci decreased less than R2 and the AMC decreased at R4 and R6. The ordinary varieties decreased significantly. This suggested that the stomatal limitation might have led to photosynthesis decline. The photosynthetic electron transfer quantum efficiency (Φ PSII) and photochemical quenching (qP) and apparent photosynthetic quantum transfer efficiency (ETR) decreased and the photochemical quenching (NPQ) increased under drought stress. The original light energy transformation efficiency (Fv/Fm) showed no apparent change. The MDA, proline and soluble sugar content increased under drought stress.
To study the changes of constituents in Camellia oleifera Abel among different growth stages, the seeds of the Changlin-4, Changlin-40 and Changlin-166 were picked from May to October. Their weight，relative amount of water, oil, protein, starch, soluble sugar and tea saponin in seeds were tested after July. The constitute of fatty acids, hydrolyzed amino acids and free amino acids were also analyzed. The results showed: 1) the seeds grew slowly before July and after that their growth rate increased until September, and their weight hardly increased any in October; 2) during the maturing of the seeds, the water content decreased constantly but the organic content increased quickly, in which the oil increased the most, and the successive were the tea saponin, protein and starch. Only the soluble sugar content was relatively stable; 3) the hydrolyzed amino acids content increased month by month and the content was different among different type of hydrolyzed amino acids. the content of Gly, Arg and Leu were the greatest and Cys were the least. However, the content of free amino acids didn’t always grow with the seeds. The rank of different free amino acids content shown was almost the same as those of the hydrolyzed amino acids; 4) the constitution of the fatty acid had a great difference among different months. The oleic acid content increased greatly with the seeds maturation, which increased to the 80% of the whole fatty acid content. The linoleic acid had a high content in July but decreased quickly with the growth of the seeds, so did the content of linolenic acid and palmitic acid. The stearic acid content was low and got little increase each month. The content of palmitoleic acid and Cis-11-eicosenoic acid was low in various periods. This article concludes that the variation of constituent content in Camellia oleifera Abel seeds was associated with different growth stages. It provided reference value to the orderly improvement of Camellia oleifera Abel quality, as well as in improving the content of useful material by molecule and physiology methods at critical period, which in turn would improve the effectiveness of Camellia oleifera Abel.
The mycelium growth rate was used to determine the toxicities of tebuconazole，prochloraz, fludioxonil and kresoxim-methyl to Sclerotium rolfsii, and field trials were conducted to investigate the control efficacy of four fungicides by seed dressing and irrigation pier in fruiting stage. Four fungicides could significantly inhibit the mycelia growth of Sclerotium rolfsii, and the EC50 values are 0.05, 0.03, 0.22 and 0.61 mg/L respectively. The control effects of irrigation pier treatments of four fungicides were better than seed dressing in field trials. The control effect of tebuconazole was relative higher, with 62.91% -74.93 % by seed dressing, and 68.60%-82.87% by irrigating pier. All treatments could strongly increase the peanut yield and the growth rare of irrigation pier treatment was higher than that of seed dressing treatment. The yield growth rate of tebuconazole treatment was the highest with 10.74%-11.55% increase.
Strongyllodes variegatus (Coleoptera: Nitidulidae, Fairmaire，1891) was first found on oilseed rape in Chaohu area, Anhui Province, China in 2008. The damage it caused on rapeseed had become more serious in recent years. Field surveys and tests showed that S. variegatus mainly distributed in Chaohu, Ma’anshan, Xuancheng, Anqing, Liu’an, (part of) Hefei and (part of) Chuzhou in Anhui Province. The pest was observed to reproduce twice a year and occurred in spring (from March to May) and autumn (from September to November) in Anhui Province. But the main plague and reproduction period were from March to May. Adults fed on rape leaves, tender stem, buds and tender pods, which led to crescent wounds on the undersurface of leaves and bare stems. The larvae only eat mesophyll between the surfaces of leaves, forming irregular bubble damage, which led to leaf rot and fall before senescence. The over-summering adults hatched their eggs in soil and fed on leaves during September to November. The elder larvae pupated and emerged in soil. The adults aestivated and hibernated in soil and among rapeseeds.
The objective of this study is to research the impact of glyphosate on yield component and resistance genetics of glyphosate-tolerant soybean and to provide a statistical basis for applying glyphosate effectively to control weed. The random complete block design was used and the influence of spraying 41% glyphosate isopropyl ammonium AS at different rates on yield component and transgenic ratio of transgenic soybean cultivar GTS 40-3-2 at V2 stage was studied in the field. The following year, the glyphasate resistance and yield component of first filial generation soybean after applying glyphosate in the same concentration rates were investigated. The result indicated that glyphosate was effective at 1.23-12.30 kg (ai) ? hm-2 to control weed. However, the glyphosate at 7.38-12.30kg (ai) ? hm-2 markedly reduced the number of seeds and seed weight per plant of soybean parents and progeny. Spraying glyphosate had no obvious impact on the quantity of genetically modified ingredient (GM %) in seeds of GTS 40-3-2,? which induced the first filial generation soybean with the same glyphosate tolerance as their parents. It was concluded that spraying glyphosate at 1.23-4.92 kg (ai) ? hm-2 was available for weeding control as well as maintaining the yield component of GTS 40-3-2, which could be safe to use at V2 stage of the glyphosate-tolerant soybean. Application of glyphosate above 7.38 kg (ai) ? hm-2 depressed the seed number but did not affect the seed mass of GTS 40-3-2. Spraying glyphosate did not have negative influence on the genetics of resistance to glyphosate of GTS 40-3-2. Glyphosate could be used to weed control with recommended rate when the first filial generation of GTS 40-3-2 was cultivate
Based on the polymerase chain reaction-denaturing gradient gel electrophoresis （PCR-DGGE) and sequence analysis, two herbicide tolerant genetically modified soybeans (PAT, ALS) and the corresponding non-transgenic soybeans (PAT1, ALS1) and the local main non-transgenic soybean (Zhonghuang 13) were selected to investigate the effects of herbicide tolerant transgenic soybean cultivation on nitrogen-fixing bacteria in rhizosphere under field experiment,. The results showed that the community composition similarity of soil nitrogen-fixing bacteria between transgenic soybean (PAT, ALS) and the corresponding non-transgenic soybean (PAT1, ALS1) was approximately 60%. Compared to the corresponding non-transgenic soybeans (PAT1, ALS1), the diversity index (H) and evenness index (EH) of nitrogen-fixing bacterial nifH genes in rhizoshpere of herbicide tolerant genetically modified soybeans (PAT, ALS) both showed no significant difference (p>0.05). The sequencing and phylogenetic analysis indicated that nitrogen-fixing bacterial nifH genes in rhizoshpere of herbicide tolerant genetically modified soybeans (PAT, ALS) and the corresponding non-transgenic soybeans (PAT1, ALS1) mainly belonged to cyanobacteria, Proteobacteria and Firmicutes. Nitrogen-fixing microbial communities were significantly (p<0.05) influenced by the levels of the available phosphorus (AP) and nitrate nitrogen (NO3--N) when canonical correspondence analysis was used to identify relationship between nifH gene and soil physicochemical factors. This result indicated that herbicide tolerant genetically modified soybeans had no significant effects on the diversity of soil nitrogen-fixing bacterial nifH genes in rhizosphere.
A novel method was developed to determine resveratrol in peanut by HPLC with on-line solid phase extraction. The on-line soild phase extraction column was prepared by packing dsDNA column filler in stainless column. The loading and washing solution of the extraction column were both water, the transferring solution was acetonitrile-water (80:20, V/V), and the determining solution was acetonitrile-water (25:75, V/V). Under the above condition, the linear regress equations was Y=1 843.805X+39.179 1, with correlation coefficients of 0.999 78, detection limits of 0.004mg/kg, and quantification limits of 0.01mg/kg. The average recoveries of 4 levels spiking were 86.80 to 105.37%, the relative standard deviation (RSD) was 0.25 to 7.05% (n=6).
An enzymatic synthesis process for a new functional lipid prepared with rutin and α-linolienic was established using commercially immobilized lipase from Candida antarctica (Novozym435) as biocatalyst. Response surface methodology (RSM) was adopted to evaluate the effects of reaction parameters on esterification rate. The optimum process parameters were as follows: 17.9mg/mL Novozym435, 17.3mmol/L rutin concentration, 1:4.5 molar ratio of rutin esters to α-linolienic, 60h for reaction in acetone, with esterification rate of 92.6%. After purification, the purity of products reached up to 95.0%.
With the aim to identify the role of salicylic acid (SA) in peanut induced resistance to Peanut web blotch, the content of endogenous SA in leaves and roots of peanut plants were measured by Enzyme-Linked Immunosorbent Assay (ELISA) method. Then the changs of SA content in leaves at different location inoculated with 2% cultural filtrate of Phoma arachidicola were monitored, and their relationship with induced resistance were analyzed. The results indicated no significant differences in content of SA in leaves at different locations without inoculation at the same growth stage, However, the content of SA in leaf was higher than in roots. After treated with 2% culture filtrate, the content of SA reached its peak twice both in treated leaves and untreated nearby leaves. At first peak, there were no significant differences in both peak time and peak values between different leaves; at second peak, both peak time and peak values were closely related to leaves in different locations. It was concluded that SA played a significant role in peanut web blotch resistance and signal transduction.
Soybean cyst nematode (SCN) is the most economically damaging pathogen of soybeans in the world. The research results show that plants inhibiting the development of SCN by secreting defense enzymes, phenolic metabolism, phytoalexins and so on, is a plant resistance mechanism. rhg1 and rhg4 allele are widely used for resistance against SCN. Pel, GmHs1pro-1, 30C02 and GmRLK18-1 are genes or protein encoded by gene for resistance against SCN. This paper reviewed the plant resistance mechanism against SCN and the research progress on resistant genes, and provided a reference for soybean molecular breeding.
