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2017 Vol. 32, No. 11

Crop Science
CRISPR/CAS9-mediated Editing for Pollen-specific OsIPA Genein Rice
YANG Shao-hua, CHEN Hui-mei, ZHOU Shu-fen
2017, 32(11): 1173-1177. doi: 10.19303/j.issn.1008-0384.2017.011.001
Abstract:
Male gamete development is an essential step in rice reproduction process, and male sterility is crucial for the utilization of rice heterosis. Since the anther or pollen specific genes in rice might play a pivotal role in the male gamete development, the functions of OsIPA were of great interest to the scientists. This study applied CRISPR/CAS9 to generate an OsIPA mutant for an investigation. Two sites in the 1st exon of OsIPA were selected as the targets. After sequencing, a total of 8 different types of mutant, including deletion, substitution and single nucleotide insertion, were found in 24T0 transgenic plants.
Joint Multi-Generation Genetic Analysis on Kernelling Rate of Maize
LI Hong-tao, XU Han-yuan, CHI Ming, HUA Guo-dong, WANG Jun
2017, 32(11): 1178-1184. doi: 10.19303/j.issn.1008-0384.2017.011.002
Abstract:
The kernelling rate of maize is an indicator of the as similation product distribution of the plant. It closely relates to the kernel count on an ear of maize, or the plant yield. To understand the inheritance of maize would be of significant interest inbreeding high yield varieties.Six generations, i.e., P1, P2, F1, BC1, BC2, and F2, from the cross L055 Qi319 were studied using the mixed major genesand ploygene inheritance model. The results showed that maize kernelling rate was controlled by two major genes with additive-dominance-epistatic effects as well as polygene with additive-dominant effects. The additive-dominant effects of the two major genes and polygene could increase the rate, while the epistatic effect decreased it.The major gene heritability of BC1, BC2, and F2 were 54.05%, 36.26% and 48.83%, respectively; while the polygene, 26.45%, 46.36% and 31.43%, respectively.The combined heritability of major genes and polygene was 81.13%.They were all important on the inheritance of kernelling rate, which was mainly governed by the major genes, affected more strongly by the non-additive than the additivegene effect, and somewhat influenced by the environmental conditions.
Effect of Various Fertilizers on Growth of Elephantopus scaber Linn Intercropped in Forest
XIE Xue-fang, ZHANG Yong-fa, FENG Ming, LI Yi-jian, ZHENG Ding-hua
2017, 32(11): 1185-1188. doi: 10.19303/j.issn.1008-0384.2017.011.003
Abstract:
Effect of various fertilizers for the intercropped Elephantopus scaber Linn in forest on the growth performance of the grass was studied. Major growth and yield indices of the turf with or without the application of a biogas fertilizer, a compound fertilizer or organic manure were compared. The results showed that the fertilization one month after planting effectively increased the plant height, leaf count, leaf length, leaf width, root length, total yield, root yield, root/shoot ratio, and other growth and yield indices of the grass. It doubled the total yield as compared to control. Consequently, fertilization could be a direct and effective way to promote the growth and yield of E. scaber. The application was best implemented from May to August when the peak growth of the grass occurred. The biogas slurry appeared to perform better overall and was more conducive to the grass-intercropping than the other fertilizers tested.
Animal Science
Efficacy Evaluation of a Concentrated Inactivated Vaccine against Novel Duckling Reovirus Infection
CHEN Shi-long, XIAO Shi-feng, CHENG Xiao-xia, LIN Feng-qing, WANG Shao, WANG Jin-xiang, ZHU Xiao-li, CHEN Shao-ying
2017, 32(11): 1189-1192. doi: 10.19303/j.issn.1008-0384.2017.011.004
Abstract:
In this paper, an experimental concentrated inactivated vaccine and a conventional inactivated vaccine against Novel Duckling Reovirus (NDRV) infection were prepared. After one single-dose immunization in adult Cherry Valley ducks, the serum samples collected in a period of 13 weeks were tested for antibodies by serum neutralisation (SN) method. The protective effects against NDRV infection were observed in 3-day-old Cherry Valley ducklings and mule duckling after a single-dose immunization. The serological results showed that concentrated inactivated vaccine was much more immunogenic than the conventional inactivated vaccine, with a much higher SN antibody titers at all test time points. The concentrated inactivated vaccine was highly protective against NDRV challenge at 7-day post-vaccination with a 100% protective efficacy, as compared to 40%-60% protective efficacy for conventional inactivated vaccine. Taken together, this experimental concentrated inactivated vaccine exhibited immune efficiency much superior to conventional inactivated vaccine and might be a useful vaccine against NDRV infection.
Establishment of One-step RT-PCR for Batai Virus Detection in Ducks
FU Guang-hua, CHEN Cui-teng, FU Qiu-ling, LIU Rong-chang, CHENG Long-fei, SHI Shao-hua, WAN Chun-he, CHEN Hong-mei, HUANG Yu
2017, 32(11): 1193-1196. doi: 10.19303/j.issn.1008-0384.2017.011.005
Abstract:
For a rapid detection of Batai virus (BATV) in ducks, a one-step reverse transcription polymerase chain reaction (RT-PCR) assay was developed based on the conserved region of the M gene in BATV. With the optimized reaction conditions, this assay could specifically amplify a 480 bp fragment from the M gene. There were no similar gene amplifications on 5 other pathogens found in ducks, such as avian Tembusu virus and duck hepatitis virus. The testing sensitivity of the method was 1×101.1 TCID50 with a high reproducibility. Consequently, this newly developed methodology seemed to be highly specific and sensitive for BATV detection, and could be applied for rapid and early diagnosis as well as epidemiological investigations on BATV.
Effects of Probiotics Fermentation Products on Meat Production and Serum Indices of Mindong Goats
CHEN Xin-zhu, ZHANG Cheng-jin, LIU Yuan, YAN Di, LI Wen-yang, WENG Bo-qi
2017, 32(11): 1197-1200. doi: 10.19303/j.issn.1008-0384.2017.011.006
Abstract:
Effects of including fermentation products of yeasts and lactic acid bacteria(LAB) in feed on the meat production and serum indices of Mindong goats were studied.Thirty-two male and 32 female goats of similar age and physical conditions were randomly and evenly divided into a treatment and a control group for a month-long feeding test.At the end, blood samples from 16 randomly picked goats in each group were collected for serum analysis.The results showed that the averaged final body weight and daily weight gain of the goats in the treatment group were significantly higher than those of control(P < 0.05).In the treatment goats, the serum albumin, globulin ratio, and alkaline phosphatase were extremely significantly lower(P < 0.01), glutamic-pyruvic transaminase, total cholesterol, high-density lipoprotein, and low-density lipoprotein significantly lower(P < 0.05), while the immune globulin and IgA extremely significantly higher(P < 0.01) than control. As tested by the enzyme-linked method, the immune globulin IgE and IgM in the treatment goats significantly increased at P < 0.01 level, while IgA significantly increased at P < 0.05 over control. It indicated that the dietetic inclusion of mixture of the yeast and LAB fermentation extracts could improve the meat production, metabolic efficiency and immunity of Mindong goats.
Horticultural Science
Selection and Validation of Reference Genes for Real-time Florescence Quantitative PCR Analysis on Gene Expression of White Tea
CHEN Jing, ZHENG Zhi-lin, LIN Yi, CAO Hong-li, CHEN Di, CHEN Gui-xin, YE Nai-xing
2017, 32(11): 1201-1206. doi: 10.19303/j.issn.1008-0384.2017.011.007
Abstract:
To select appropriate reference genes to be used in the real-time florescence quantitative PCR analysis for studying the gene expression in of white tea leaves during withering, the software of GeNorm, NormFinder, and BestKeeper were applied. Three stable reference genes, β-Actin, GAPDH and RUBP, were identified. Among them, RUBP was found most adequate for comparing the expressions at various stages of withering, while β-Actin for differentiating them in leaves from different parts of a tea plant. The combination of GAPDH+RUBP was determined as choice reference genes for the analysis.
Concentrations and Distributions of Iron and Manganese in Parts of Tea Plant
SHAN Rui-yang, CHEN Yu-zhen, WANG Feng, LIN Dong-liang, ZANG Chun-rong, GAO Jun-jie, CHEN Chang-song, YOU Zhi-ming, YU Wen-quan
2017, 32(11): 1207-1212. doi: 10.19303/j.issn.1008-0384.2017.011.008
Abstract:
Inductively coupled plasma mass spectrometry was applied to determine the contents of iron and manganese in teas from Fujian province. The distribution and accumulation characteristics in different parts of a tea plant were analyzed. The results showed that the mineral concentrations significantly differed among the varieties of teas studied. The iron contents ranged from 73.36 mg·kg-1 to 284.85 mg·kg-1 with a mean of 125.60 mg·kg-1, and manganese from 201.86 mg·kg-1 to 4 905.78 mg·kg-1 with a mean of 1 651.24 mg·kg-1. Overall, the black and the green teas contained more iron than oolong tea, whereas, oolong tea had a higher manganese content than the green or the black tea. There was no significantly difference on manganese content found between the green and the black tea. Significantly differences in the mineral contents were also shown among different parts of a tea plant. The iron concentrations in different parts ranked in the order of the fine roots (6 265.07 mg·kg-1) > taproots (3 250.77 mg·kg-1) > main stem (2 178.56 mg·kg-1) > stem side (1 025.81 mg·kg-1) > old leaves (347.50 mg·kg-1) > young leaves (135.70 mg·kg-1). Thus, most of the iron was absorbed and accumulated in the roots and stems on a tea plant. On the other hand, the manganese in old leaves was the highest (P < 0.05), and evenly distributed in other parts throughout the plant. In general, Tieguanyin and Meizhan teas had greater iron or manganese contents as well as on enrichment coefficients in different parts of a tea plant than Jinguanyin or Jinxuan.
Codon Usage Bias of Chalcone Synthase Gene CHS in Kiwifruit
CHEN Yi-ting, LAI Rui-lian, FENG Xin, GAO Min-xia, ZHANG Chang-he, WU Ru-jian
2017, 32(11): 1213-1217. doi: 10.19303/j.issn.1008-0384.2017.011.009
Abstract:
To investage codon usage characteristic of chalcone synthase gene of kiwifruit, CodonW, SPSS, MEGA software and EMBOSS online program were used to analyse codon usgae bias (CUB) of CHS, and phylogenetic tree of CHS between monocotyledon and dicotyledon was constructed in this paper. The results showed that, the effective number of codons (ENc), total GC content (GC) and GC content on the 3rd site (GC3s) was 57.21, 0.533 and 0.607, respectively. The number of condons, the number of relative synonymous codon usage (RSCU) of which was more than 1.0, was 30, but no codons was with strongly CUB. The cluster tree of CHS based on CUB was closer to the real category system than it did by sequence similarity. According to codon usage frequency, the CUB between CHS of kiwifruit and Arabidopsis thaliana, Escherichia coli were more similar. These results indicated that, the CUB level of CHS was low in kiwifruit, but it was bias toward GC rich codons and synonymous codons with G or C at 3rd codon position. And CHS in monocotyledons and dicotyledons had the distinctive CUB rules. In addition, Arabidopsis thaliana and Escherichia coli migth be the best receptors for genetic transformation and heterologous expression of CHS in kiwifruit.
Bioinformatics and Identification of eIF5A Gene Family in Phalaenopsis equestris (Schauer) Rchb. f.
ZHANG Jie-wei, TANG Jiu-yang, CHEN Lan-fen, WANG Ning-lin, WANG Hong-zhi, WEI Jian-hua
2017, 32(11): 1218-1223. doi: 10.19303/j.issn.1008-0384.2017.011.010
Abstract:
To understand the functions of eIF5A gene family in Phalaenopsis equestris, the genome database and bioinformatics were applied to obtain their gene structures, coded proteins, and predicted phosphorylation sites for analysis. The results showed that the eIF5A1 and eIF5A2 genes in P. equestris genome contained 5 and 6 extrons, respectively. The multiple alignments and motif displays rendered from the MEME method indicated that all PeeIF5A1 and PeeIF5A2 proteins contained one conserved DNA binding OB binding motif, PF01287. The phosphorylation site prediction on the proteins showed numerous sites.These results would provide the important clues for functional studies to reveal the role of PeeIF5A1 and PeeIF5A2 proteins in P.equestris.
In Vitro Propagation of Dan-Shui Pear
ZENG Ling-da, LI Cai-hua, ZHAI Qiu-yan
2017, 32(11): 1224-1227. doi: 10.19303/j.issn.1008-0384.2017.011.011
Abstract:
To establish an in vitro culture system for propagating Dan-Shui pear, Xiangshui pear was used on Murashige and Skoog medium (MS) as the basal medium for the experimentation. Conditions of the tissue culture, such as hormone combination and concentration on explant induction and proliferation culture, as well as hormone types and concentrations on rooting culture, were studied. The optimal combination for the axillary bud induction was found to be MS+ 6-BA 1.0 mg·L-1+NAA 0.1 mg·L-1 that yielded a germination rate of 71.7%. For the bud proliferation, it was MS+6-BA 3.0 mg·L-1+NAA 0.20 mg·L-1 to deliver a multiplication coefficient of 3.2. For the optimal rooting performance, the combination of 1/2 MS+IBA 0.3 mg·L-1 to result in a rooting rate of 63.3% was selected. Thus, addition of the growth regulators to MS medium benefitted the in vitro propagation of Dan-Shui pear.
Effects of Types of Covering Pouch on Quality of Jumeigui Grapes
LIU Xin-ming, XU Lu-jie, CHEN Ting, WANG Jian-chao, ZHU Su-fang, LEI Yan
2017, 32(11): 1228-1233. doi: 10.19303/j.issn.1008-0384.2017.011.012
Abstract:
Effect of covering pouches of different materials on quality of the bagged grapes was studied. Pouches made of white, wood pulp paper, translucent plastic film, and nylon mesh were used to cover Jumeigui grapes on the vines 15 ds after flowering. Regardless the differences in material, the bagging increased individual fruit weight and soluble solids content, and reduced titratable acid as compared to control with full exposure to the ambient conditions. The white paper pouch showed the most desirable results over all, as the fully ripened grapes had an appealing shape with a hardness of 0.081 kg·cm-2, weight at 10.27 g·fruit-1, soluble solids at 19.00%, total acid at 0.208%, Vitamin C at 4.410 mg·hg-1, solids/acid ratio of 91.35, and stability for shipping and storage. The result provided a positive direction for the standardized production of high quality grapes in the southern regions.
Utilization of Soybean Strawsfor Cultivating Lentinula edodes
ZHANG Yu-mei, HU Run-fang, CHEN Wei, CHEN Yu-hua, LIN Guo-qing
2017, 32(11): 1234-1238. doi: 10.19303/j.issn.1008-0384.2017.011.013
Abstract:
Differently formulated substrates consisting of soybean straws to replace sawdust or wheat bran were used to cultivate Lentinula edodes for evaluation. The resulting yield, economics, amino acid content, and production cost of the mushroom fruit bodies were analyzed. It was found that the medium containing 15% of the soybean straws could be applicable for the cultivation. In fact, as high as 35% of soybean straws in a medium could substitute wheat branto achieve a same fresh mushroom production and amino acid content in the fruiting bodies. By doing so, a saving of 4 250 yuan in producing 10 000 bags of L. edodes could be realized. The information suggested means to effectively utilize soybean straws for the mushroom cultivation.
Standards for Grading Anoectochilus roxburghii (Wall.) Lindl.Seedings
ZHAO Yun-qing, CHEN Jing-ying, HUANG Rui-ping, HUANG Ying-zhen
2017, 32(11): 1239-1243. doi: 10.19303/j.issn.1008-0384.2017.011.014
Abstract:
Standards for grading seedlings of Anoectochilus roxburghii (Wall.) Lindl. were studied. Factors includingplant height and weight, average length, width and count of leaves, stem diameter, andinternode countof a seedlingpropagated from tissues were analyzes on their interrelations and chemical compositions to select agrading system.Cluster and statisticalanalyses were performed to weigh those factors against the survival rate of the seedlings. The results indicated that the plant height and weight, as well as the stem diameter were the determining indicators for an effective grading. Three classifications were thus established:the 1st Grade to show a seedling height of ≥ 130 mm, a stem diameter of ≥ 2.8 mm, and a weight of ≥ 1.3 g; the 2nd Grade, a seedling height of ≥ 110 mm, a stem diameter of ≥ 2.4 mm, and a weight of ≥ 1.0 g; and, the 3rd Grade, a seedling height of ≥ 90 mm, a stem diameter of ≥ 2.1 mm, and a weight of ≥ 0.6 g.The proposed standards would allow a clearly definedevaluation on seedings in the breeding practices.
Plant Protection
Cloning and Expression of Bursicon Gene of Myzus persicae (Sulzer)
ZHANG Chao, YANG Hong, YANG Mao-fa, YU Hui-ping Wei, ZHANG Wei
2017, 32(11): 1244-1250. doi: 10.19303/j.issn.1008-0384.2017.011.015
Abstract:
Bursicon regulates the epidermal sclerosis and wing-spreading in insects. It plays a key role in epidermis hardening of insects after molting. The full-length sequences of the bursicon genes (burs-α and burs-β) of M. persicae (Sulzer) were obtained through cloning for this study, and named Mpburs-α (GenBank accession number:MF083566) and Mpburs-β (GenBank accession number:MF083567), respectively. Mpburs-α contained a 483 bp open reading frame (ORF) and was encoded 160 amino acid residues. Its molecular mass was predicted to be 17.59 kDa with a formula of C759H1204N210O232S19 and an isoelectric point of 7.96. Mpburs-β contained a 417 bp ORF and was encoded 138 amino acids. Its molecular mass to be 15.45 kDa with a formula of C672H1072N180O212S12 and an isoelectric point of 4.8. The phylogenetic tree analysis showed a relatively close genetic relationship of M. persicae with Acyrthosiphon pisum (Harris) and Diuraphis noxia (Mordvilko). The qRT-PCR results indicated that Mpburs-α and Mpburs-β expressed during the entire developmental stage of M. persicae and arrived at a peak expression level in the 1st instar nymph. The expression levels of the two genes in the winged adult aphids were significantly higher than those in the wingless adult aphids suggesting an important role played by bursicon in the wing development for the insect. The finding might lead to further studies on the functions of bursicon in the molting epidermal ossification and wing formation of M. persicae.
Effects of Processing and Temperature on Imidacloprid Residues in Chrysanthemum morifolium
LU Yan-ping, YANG Guang, WU Jia-lun
2017, 32(11): 1251-1256. doi: 10.19303/j.issn.1008-0384.2017.011.016
Abstract:
Effects of processing and temperature on the residual content of imidacloprid in Chrysanthemum morifolium were investigated. As indicated by the level of the pesticide residues in commercial C. morifolium products on the market, the common field spraying practice on the plants seemed acceptable for meeting the food safety requirement. For this study, at various processing stages the imidacloprid concentrations in the dried chrysanthemum buds and flowers were reduced by 6.36% and 8.71%, respectively, during blanching; 3.99% and 4.48%, respectively, during thermal dehydration; and, 13.18% and 29.60%, respectively, during ambient temperature drying. It showed that imidacloprid was largely dissipated from the products by drying under ambient conditions. Evaporation apparently raised the relative imidacloprid concentration in the finished products. Under high temperatures, the rate of imidacloprid reduction in a thermal dehydration was greater than that of an ambient drying suggesting the possibility of pesticide degradation by heat.
Resources and Environmental Science
Molecular Identification and Productivity on Hyaluronic Acid in Submerged Culture of Different Cordyceps cicadae Strains
LONG Liang-kun, HUANG Shu-hui, SHI Zi-yu, LIN Qun-ying, LI Chuan-hua, DING Shao-jun
2017, 32(11): 1257-1262. doi: 10.19303/j.issn.1008-0384.2017.011.017
Abstract:
This study aimed to identify 4 Cordyceps cicadae strains and compare their productivities on hyaluronic acid (HA) in a liquid culture. The molecular identification for CC1502, CC1504, CC1601 and CC1602 was conducted based on their internal transcribed spacer (ITS) sequences and using the randomly amplified polymorphic DNA (RAPD) analysis. The results indicated that there was no difference on ITS sequences among these fungal strains, and that the RAPD patterns were amplified by the primer, AP-A20, AP-D18 or AP-H18. As shown by the dendrogram analysis with UPGMA method, CC1504 had a low genetic similarity with any of the other strains, and CC1601 related closely to CC1602. The liquid fermentation of the strains showed different levels of HA productivity between CC1502 and CC1504, and glucose and peptone were the choice carbon and nitrogen sources. The maximum HA production was up to 2.45 mg·mL-1 generated by CC1502, which, therefore, was considered a candidate for industrial application in the future.
Food Science
Tablet-forming Process for Utilization of Substandard Okra
WENG Min-jie, CHEN Jun-chen, LAI Pu-fu, TANG Bao-sha, SHEN Heng-sheng
2017, 32(11): 1263-1268. doi: 10.19303/j.issn.1008-0384.2017.011.018
Abstract:
To maximize the utilization of okra resources, a process was developed to make nutritional tablets from substandard material that would be otherwise of low market value or discarded as waste. The formulation and process for the tablet formation were optimized by using the response surface methodology. Disintegration time and friability of the tablets were the criteria for the evaluation under three process parameters including additions of sodium carboxymethyl starch and microcrystalline cellulose in the formulation as well as pressure applied to compress and form the tablets. The optimized conditions were (a) a mixture of 76.6% okra powder, 12.6% of sodium carboxymethyl starch, and 10.8% of microcrystalline cellulose, and (b) 17.6 MPa pressure for the compression. The tablets obtained had a friability of 0.308% and a disintegration time of 1 180 s, with a perfect shape, smooth surface and high palatability. It appeared that the experimental results could be use for scale up to manufacture a functional food product from the substandard okras.
Analysis and Evaluation of Nutritional Components of Brown species of Auricularia polytricha
LIN Xiang-xin, YAN Sun-an, YAO Qing-hua, CHEN Guo-ping, LIN Qiu
2017, 32(11): 1269-1274. doi: 10.19303/j.issn.1008-0384.2017.011.019
Abstract:
Auricularia polytricha is an economically important mushroom species in southern China. Nutritional compositions were compared between normal color and brown color variant groups. According to national standards, nutrition in brown species and normal species of Auricularia polytricha were compared. Results showed that the contents of protein, reducing sugar, vitamin B1 and vitamin B2 of Auricularia polytricha is significantly lower than brown species. The contents of E/P value and Fe of Auricularia polytricha is significantly higher than brown species. But results on fatty, crude ash, dietary fiber, energy maintained similar between them. Both of them contained more than 18 kinds of amino acid. The content of each amino acid, essential amino acids, no-essential amino acids, essential amino acids for child, all amino acids, sweet amino acids, acidic amino acids, bitter amino acids, flavor amino acids, sulfur amino acids, branched-chain amino acids and aromatic amino acids in brown species is higher than normal species, but E/T、E/N is lower than normal species. The nutritional value of AAS, CS and SRCAA in brown species is higher than normal species. It indicated that significant differences were observed on nutrition traits between these two groups and the nutrition of amino acids in brown species was better than normal species.