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Dietary aflatoxin B1 induces abnormal deposition of melanin in the corium layer of the chicken shank possibly via promoting the expression of melanin synthesis-related genes

WANG Yong-li, HUANG Chao, YU Yang, CAI Ri-chun, SU Yong-chun, CHEN Zhi-wu, ZHENG Maiqing, CUI Huan-xian
2023, 22 (6): 1847-1856.   DOI: 10.1016/j.jia.2023.04.001
Abstract232)      PDF in ScienceDirect      

San-Huang chicken is a high-quality breed in China with yellow feather, claw and break.  However, the abnormal phenomenon of the yellow shank turning into green shank of San-Huang chicken has been a concern, as it seriously reduces the carcass quality and economic benefit of yellow-feathered broilers.  In this study, the cause of this abnormal green skin in shank was systematically investigated.  Physiological anatomy revealed that the abnormal skin in shank was primarily due to the deposition of melanin under the dermis.  After analyzing multiple potential causes such as heredity (pedigree and genetic markers), environment (water quality monitoring) and feed composition (mycotoxin detection), excessive aflatoxin B1 (AFB1) in feed was screened, accompanied with a higher L-dihydroxy-phenylalanine (L-DOPA) (P<0.05) and melanin content (P<0.01).  So it was speculated that excessive AFB1 might be the main cause of abnormal green skin in shank.  Subsequently, the further results showed that a high concentration of AFB1 (>170 μg kg–1) indeed induced the abnormal green skin in shank compared to the normal AFB1 content (<10 μg kg–1), and the mRNA levels of TYR, TYRP1, MITE, MC1R and EDN3 genes related to melanin deposition would significantly up-regulate (P<0.01) and the content and activity of tyrosinase (TyR) significantly increased (P<0.05).  At the same time, the content of L-DOPA and melanin deposition also increased significantly (P<0.01), which also confirmed the effect of excessive AFB1 on melanin deposition in skin of shank.  Results of additional experiments revealed that the AFB1’s negative effect on melanin deposition in skin of shank could last for a longer time.  Taken together, the results of this study explained the occurrence and possible mechanisms of the abnormal AFB1-related green skin in shank of chickens.  Excessive AFB1 in diets increased the L-DOPA content and melanin abnormal deposition in the chicken shank possibly via promoting TyR content and activity, and the expression of melanin synthesis-related genes.  Furthermore, our findings once again raised the alarm of the danger of AFB1 in the broiler production.

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Effect of dioscorea opposite waste on growth performance, blood parameters, rumen fermentation and rumen bacterial community in weaned lambs
GUO Yun-xia, YANG Ruo-chen, DUAN Chun-hui, WANG Yong, HAO Qing-hong, JI Shou-kun, YAN Hui, ZHANG Ying-jie, LIU Yue-qin
2023, 22 (6): 1833-1846.   DOI: 10.1016/j.jia.2022.10.002
Abstract293)      PDF in ScienceDirect      

This study investigated the effects of dioscorea opposite waste (DOW) on the growth performance, blood parameters, rumen fermentation and rumen microbiota of weaned lambs.  Sixty healthy weaned Small-Tailed Han lambs (male, (22.68±2.56) kg initially) were used as the experimental animals.  Four levels of concentrate: 0 (control, CON), 10% (DOW1), 15% (DOW2) and 20% (DOW3), were replaced with DOW in the basal diet as experimental treatments.  The results showed that lambs fed the DOW2 diet had a higher (P<0.05) dry matter intake (DMI) than the other groups.  There was no significant difference (P>0.05) among DOW groups in average daily weight gain (ADG), and replacing concentrate with DOW linearly or quadratically increased (P<0.05) the ADG, while lambs fed the DOW2 diet showed greater (P<0.05) ADG than the CON group.  The relative plasma concentration of growth hormone (GH), insulin like growth factor-1 (IGF-1) and insulin were affected by DOW, replacing concentrate with DOW linearly or quadratically (P<0.05) enhanced the plasma concentration of GH, IGF-1 and insulin, which was significantly higher (P<0.05) in the DOW2 group than in the CON, DOW1 and DOW3 groups.  In addition, the DOW treatment showed a lower (P<0.05) concentration of blood urea nitrogen (BUN) than the CON group.  Replacing concentrate with DOW quadratically decreased (P<0.05) the ruminal ammonia nitrogen (NH3-N) and increased (P<0.05) the total of volatile fatty acids (TVFAs) at 0 and 4 h after feeding as well as linearly decreased (P<0.05) the NH3-N at 8 h after feeding.  Replacing concentrate with DOW linearly decreased (P<0.05) the propionate and increased the aceate before feeding, and linearly decreased (P<0.05) propionate and quadratically increased (P<0.05) the aceate at 4 and 8 h after feeding.  Lambs fed the DOW2 diet increased the phylum Firmicutes and genera Succiniclasticum and Ruminococcus_1 groups, whereas decreased (P<0.05) the relative abundance of phylum Deferribacteres and genera intestinimonas and Ruminiclostridium.  In summary, replacing the concentrate with 15% DOW was beneficial for improving the rumen fermentation and ADG by increasing the DMI and modulating the rumen microbial community.

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Development and application of KASP marker for high throughput detection of the seedless trait in grapevine
WANG Fu-qiang, BIAN Lu, QIU Peng-peng, GUO Shuo, GUO Jing-han, GUO Chen-shuo, JIANG Jian-fu, LIU Chong-huai, WANG Yong, LIU Guo-tian, WANG Yue-jin, XU Yan
2023, 22 (11): 3269-3283.   DOI: 10.1016/j.jia.2023.10.014
Abstract207)      PDF in ScienceDirect      

Molecular marker-assisted selection (MAS) can significantly accelerate and improve the efficiency of the breeding process in seedless grape cultivars.  In this study, we developed the KASP_VviAGL11 and VviAGL11_410 markers based on a single nucleotide polymorphism (SNP) site (Chr18: 26889437 (A/C)) of the VviAGL11 gene, and compared them with previously reported SSR markers p3_VvAGL11 and 5U_VviAGL11 by testing 101 cultivars and 81 F1 hybrid progenies.  The results showed that both of the proposed markers obtained 100% accuracy rates in detecting allele A, which was closely associated with the seedless trait in grapes, while p3_VvAGL11 and 5U_VviAGL11 had lower accuracy rates due to their tendency to produce false positives.  After careful evaluation of the technical advantages and disadvantages associated with these markers, we concluded that KASP_VviAGL11 was superior in terms of simplicity, cost-effectiveness, efficiency, and accuracy.  Thus, we optimized the process of molecular MAS for seedless grapes, focusing on the KASP_VviAGL11 marker as a central component, to provide key technical support for the development of new seedless grape cultivars.

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Genome-wide association study for numbers of vertebrae in Dezhou donkey population reveals new candidate genes
SUN Yan, LI Yu-hua, ZHAO Chang-heng, TENG Jun, WANG Yong-hui , WANG Tian-qi, SHI Xiao-yuan, LIU Zi-wen, LI Hai-jing, WANG Ji-jing, WANG Wen-wen, NING Chao, WANG Chang-fa, ZHANG Qin
2023, 22 (10): 3159-3169.   DOI: 10.1016/j.jia.2023.04.038
Abstract285)      PDF in ScienceDirect      

Numbers of vertebrae is an important economic trait associated with body size and meat productivity in animals.  However, the genetic basis of vertebrae number in donkey remains to be well understood.  The aim of this study was to identify candidate genes affecting the number of thoracic (TVn) and the number of lumbar vertebrae (LVn) in Dezhou donkey.  A genome-wide association study was conducted using whole genome sequence data imputed from low-coverage genome sequencing.  For TVn, we identified 38 genome-wide significant and 64 suggestive SNPs, which relate to 7 genes (NLGN1, DCC, SLC26A7, TOX, WNT7A, LOC123286078, and LOC123280142).  For LVn, we identified 9 genome-wide significant and 38 suggestive SNPs, which relate to 8 genes (GABBR2, FBXO4, LOC123277146, LOC123277359, BMP7, B3GAT1, EML2, and LRP5).  The genes involve in the Wnt and TGF-β signaling pathways and may play an important role in embryonic development or bone formation and could be good candidate genes for TVn and LVn.

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Potential regulation of linoleic acid and volatile organic compound contents in meat of chickens by PLCD1
LIU Xiao-jing, WANG Yong-li, LIU Li, LIU Lu, ZHAO Gui-ping, WEN Jie, JIA Ya-xiong, CUI Huan-xian
2023, 22 (1): 222-234.   DOI: 10.1016/j.jia.2022.08.063
Abstract217)      PDF in ScienceDirect      

Omega-3 (linolenic acid (ALA), docosapentaenoic acid, eicosapentaenoic acid) and omega-6 (linoleic acid (LA), arachidonic acid) polyunsaturated fatty acids are essential for health and normal physiological functioning in humans.  Here we report a genome-wide association study (GWAS) on LA content in chicken meat.  The 19 significant single nucleotide polymorphisms (SNPs) identified by the GWAS approach were annotated in VILL, PLCD1 and OXSR1 genes with highly polymorphic linkage blocks, and explained 4.5% of the phenotypic variation in the LA content.  Specifically, the PLCD1 mRNA expression level was negatively correlated with the LA content, and significantly higher in chickens with low LA content than in those with high LA content.  In addition, PLCD1 was found to be involved in metabolic pathways, etc.  Furthermore, the LA content was correlated with volatile organic compounds (e.g., octanal, etc.), but no relationship was found with intramuscular fat and triglycerides in chicken meat.  The results indicated that there are key SNPs in PLCD1 that regulate the content of LA, and it has no significant effect on fat deposition, but may affect the content of volatile organic compounds (VOCs).

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Effect of the L-D1 alleles on leaf morphology, canopy structure and photosynthetic productivity in upland cotton (Gossypium hirsutum L.)
JIANG Hui, GAO Ming-wei, CHEN Ying, ZHANG Chao, WANG Jia-bao, CHAI Qi-chao, WANG Yong-cui, ZHENG Jin-xiu, WANG Xiu-li, ZHAO Jun-sheng
2023, 22 (1): 108-119.   DOI: 10.1016/j.jia.2022.08.099
Abstract170)      PDF in ScienceDirect      

One of the most important objectives for breeders is to develop high-yield cultivars.  The increase in crop yield has met with bottlenecks after the first green revolution, and more recent efforts have been focusing on achieving high photosynthetic efficiency traits in order to enhance the yield.  Leaf shape is a significant agronomic trait of upland cotton that affects plant and canopy architecture, yield, and other production attributes.  The major leaf shape types, including normal, sub-okra, okra, and super-okra, with varying levels of lobe severity, are controlled by a multiple allelic series of the D-genome locus L-D1.  To analyze the effects of L-D1 alleles on leaf morphology, photosynthetic related traits and yield of cotton, two sets of near isogenic lines (NILs) with different alleles were constructed in Lumianyan 22 (LMY22) and Lumianyan 28 (LMY28) backgrounds.  The analysis of morphological parameters and the results of virus-induced gene silencing (VIGS) showed that the regulation of leaf shape by L-D1 alleles was similar to a gene-dosage effect.  Compared with the normal leaf, deeper lobes of the sub-okra leaf improved plant canopy structure by decreasing the leaf area index (LAI) and increasing the light transmittance rate (LTR), and the mid-range LAI of sub-okra leaf also guaranteed the accumulation of cotton biomass.  Although the chlorophyll content (SPAD) of sub-okra leaf was lower than those of the other two leaf shapes, the net photosynthetic rate (Pn) of sub-okra leaf was higher than those of okra leaf and normal leaf at most stages.  Thus, the improvements in canopy structure, as well as photosynthetic and physiological characteristics, contributed to optimizing the light environment, thereby increasing the total biomass and yield in the lines with a sub-okra leaf shape.  Our results suggest that the sub-okra leaf may have practical application in cultivating varieties, and could enhance sustainable and profitable cotton production.

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Genome-wide identification of WOX gene family in apple and a functional analysis of MdWOX4b during adventitious root formation
XU Xiao-zhao, CHE Qin-qin, CHENG Chen-xia, YUAN Yong-bing, WANG Yong-zhang
2022, 21 (5): 1332-1345.   DOI: 10.1016/S2095-3119(21)63768-1
Abstract539)      PDF in ScienceDirect      
The plant-specific WUSCHEL-related homeobox (WOX) genes are crucial for plant growth and development.  Here, we systematically identified the MdWOX gene family in apple at the genome-wide level, and analyzed the phylogenetic relationships, conserved motifs, gene structure, and syntenic relationships of the MdWOX genes.  A total of 18 MdWOX genes were identified and phylogenetic analysis placed them into three clades.  The phylogenetic relationships among the WOXs were further supported by the analyses of gene structure and conserved motifs.  Chromosomal distribution and synteny analysis revealed that whole-genome and segmental duplications have played key roles in MdWOX gene family expansion.  Moreover, the MdWOX genes exhibit tissue-specific expression patterns and MdWOX4a, MdWOX4b, MdWOX5b, MdWOX11/12a, and MdWOX11/12b may play essential roles in adventitious root development.  The adventitious rooting ability was enhanced in MdWOX4b transgenic tobacco lines.  The results of this study provide useful information for future functional studies on MdWOXs in the development of apple rootstocks.  
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Analysis of combining ability for stem-related traits and its correlations with lodging resistance heterosis in hybrid wheat
YANG Wei-bing, QIN Zhi-lie, SUN Hui, HOU Qi-ling, GAO Jian-gang, CHEN Xian-chao, ZHANG Li-ping, WANG Yong-bo, ZHAO Chang-ping, ZHANG Feng-ting
2022, 21 (1): 26-35.   DOI: 10.1016/S2095-3119(20)63408-6
Abstract243)      PDF in ScienceDirect      
With the application of hybrid wheat, lodging is becoming one of the major factors limiting high yield in its production.  However, few studies have focused on combining ability and heterosis analysis of stem-related traits.  In this study, 24 crosses were made according to NCII genetic design, using the three (photo-sensitive male sterile lines)×eight (restorer lines) incomplete diallel crosses.  The length of basal second internode (LBSI) and breaking strength of basal second internode (BSBSI) as well as other stem-related traits were used to perform the principal component analysis (PCA), combining ability and heterosis analysis.  The PCA results showed that the variables could be classified into two main factors, which were named as the positive factor (factor 1) and the negative factor (factor 2), and accounted for 52.3 and 33.2%, respectively, of the total variance in different variables, combined with the analysis for index weight indicated that the factor 1-related traits play positive roles in lodging resistance formation of hybrids.  Combining ability variance analysis indicated that its genetic performance was mainly dominated by additive gene effects, and the hybrid combinations with higher lodging resistance can be selected by using of 14GF6085 (R1), 14GF6343-2 (R4), 14GF6937 (R6), 14GF7433-1 (R7), and BS1086 (M3), which are with the features with lower general combining ability (GCA) effects of factor 2-related traits whereas higher GCA effects of factor 1-related traits.  The heterosis analysis showed that the wide range of heterosis varied with the traits and combinations, and GCA or specific combining ability (SCA) effects of factor 1-related traits except wall thickness of basal second internode (WTBSI) were positively and closely related to the heterosis of lodging resistance.  Generally, the correlation coefficients of heterosis to GCA effects of sterile lines (GCAm) of factor 1-related traits are significantly higher than that to GCA of restorer lines (GCAr) and SCA, combined with the higher GCAm variance values of factor 1-related traits compared to GCAr, the GCAm of factor 1-related traits should be particularly considered when breeding hybrid combinations.  The heritability analysis showed that the narrow-sense heritability of the diameter of basal second internode (DBSI) and the center of gravity height (TCGH) were obviously lower (<60%) than other traits, suggesting that these two traits were suitable for selection in higher generation for parental breeding.  These could provide a theoretical basis for parental breeding and heterosis utilization of lodging resistance. 

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Increasing photosynthetic performance and post-silking N uptake by moderate decreasing leaf source of maize under high planting density
CAO Yu-jun, WANG Li-chun, GU Wan-rong, WANG Yong-jun, ZHANG Jun-hua
2021, 20 (2): 494-510.   DOI: 10.1016/S2095-3119(20)63378-0
Abstract103)      PDF in ScienceDirect      
To date, little attention has been paid to the effects of leaf source reduction on photosynthetic matter production, root function and post-silking N uptake characteristics at different planting densities. In a 2-year field experiment, Xianyu 335, a widely released hybrid in China, was planted at 60 000 plants ha–1 (conventional planting density, CD) and 90 000 plants ha–1 (high planting density, HD), respectively. Until all the filaments protruded from the ear, at which point the plants were subjected to the removal of 1/2 (T1), 1/3 (T2) and 1/4 (T3) each leaf length per plant, no leaf removal served as the control (CK). We evaluated the leaf source reduction on canopy photosynthetic matter production and N accumulation of different planting densities. Under CD, decreasing leaf source markedly decreased photosynthetic rate (Pn), effective quantum yield of photosystem II (ΦPSII) and the maximal efficiency of photosystem II photochemistry (Fv/Fm) at grain filling stage, reduced post-silking dry matter accumulation, harvest index (HI), and the yield. Compared with the CK, the 2-year average yields of T1, T2 and T3 treatments decreased by 35.4, 23.8 and 8.3%, respectively. Meanwhile, decreasing leaf source reduced the root bleeding sap intensity, the content of soluble sugar in the bleeding sap, post-silking N uptake, and N accumulation in grain. The grain N accumulation in T1, T2 and T3 decreased by 26.7, 16.5 and 12.8% compared with CK, respectively. Under HD, compared to other treatments, excising T3 markedly improved the leaf Pn, ΦPSII and Fv/Fm at late-grain filling stage, increased the post-silking dry matter accumulation, HI and the grain yield. The yield of T3 was 9.2, 35.7 and 20.1% higher than that of CK, T1 and T2 on average, respectively. The T3 treatment also increased the root bleeding sap intensity, the content of soluble sugar in the bleeding sap and post-silking N uptake and N accumulation in grain. Compared with CK, T1 and T2 treatments, the grain N accumulation in T3 increased by 13.1, 40.9 and 25.2% on average, respectively. In addition, under the same source reduction treatment, the maize yield of HD was significantly higher than that of CD. Therefore, planting density should be increased in maize production for higher grain yield. Under HD, moderate decreasing leaf source improved photosynthetic performance and increased the post-silking dry matter accumulation and HI, and thus the grain yield. In addition, the improvement of photosynthetic performance improved the root function and promoted post-silking N uptake, which led to the increase of N accumulation in grain.
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Drip irrigation incorporating water conservation measures: Effects on soil water–nitrogen utilization, root traits and grain production of spring maize in semi-arid areas
WU Yang, BIAN Shao-feng, LIU Zhi-ming, WANG Li-chun, WANG Yong-jun, XU Wen-hua, ZHOU Yu
2021, 20 (12): 3127-3142.   DOI: 10.1016/S2095-3119(20)63314-7
Abstract252)      PDF in ScienceDirect      
The Northeast Plain is the largest maize production area in China, and drip irrigation has recently been proposed to cope with the effects of frequent droughts and to improve water use efficiency (WUE).  In order to develop an efficient and environmentally friendly irrigation system, drip irrigation experiments were conducted in 2016–2018 incorporating different soil water conservation measures as follows: (1) drip irrigation under plastic film mulch (PI), (2) drip irrigation under biodegradable film mulch (BI), (3) drip irrigation incorporating straw returning (SI), and (4) drip irrigation with the tape buried at a shallow soil depth (OI); with furrow irrigation (FI) used as the control.  The results showed that PI and BI gave the highest maize yield, as well as the highest WUE and nitrogen use efficiency (NUE) because of the higher root length density (RLD) and better heat conditions during the vegetative stage.  But compared with BI, PI consumed more soil water in the 20–60 and 60–100 cm soil layers, and accelerated the progress of root and leaf senescence due to a larger root system in the top 0–20 cm soil layer and a higher soil temperature during the reproductive stage.  SI was effective in improving soil water and nitrate contents, and promoted RLD in deeper soil layers, thereby maintaining higher physiological activity during the reproductive stage.  FI resulted in higher nitrate levels in the deep 60–100 cm soil layer, which increased the risk of nitrogen losses by leaching compared with the drip irrigation treatments.  RLD in the 0–20 cm soil layer was highly positively correlated with yield, WUE and NUE (P<0.001), but it was negatively correlated with root nitrogen use efficiency (NRE) (P<0.05), and the correlation was weaker in deeper soil layers.  We concluded that BI had advantages in water–nitrogen utilization and yield stability response to drought stress, and thus is recommended for environmentally friendly and sustainable maize production in Northeast China.
 
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Endogenous phytohormones and the expression of flowering genes synergistically induce flowering in loquat
CHI Zhuo-heng, WANG Yong-qing, DENG Qun-xian, ZHANG Hui, PAN Cui-ping, YANG Zhi-wu
2020, 19 (9): 2247-2256.   DOI: 10.1016/S2095-3119(20)63246-4
Abstract119)      PDF in ScienceDirect      
Flowering is an important process for the reproduction of higher plants.  Up to this point, the studies on flowering have mostly focused on the model plant Arabidopsis thaliana, and the flowering mechanism of fruit trees remains mostly unknown.  The diversity of the flowering time of loquat (Eriobotrya japonica Lindl.) makes it an ideal material to study the regulation of flowering.  In this study, we first observed the inflorescence bud differentiation in two varieties of loquat that had different blooming times (cv. Dawuxing (E. japonica), that blooms in the fall and cv. Chunhua (E. japonica×Eriobotrya bengalensis Hook. f.) that blooms in the spring) and found that the starting time of inflorescence bud differentiation and the speed of inflorescence development were responsible for the difference in blooming times.  The determination of endogenous phytohormones by high performance liquid chromatography (HPLC) indicated that abscisic acid (ABA), zeatin (ZT), and gibberellin (GA3) promoted flowering in loquat, while indole-3-acetic acid (IAA) was mainly involved in inflorescence bud differentiation in Chunhua.  A transcription level analysis illustrated that multiple flowering-related genes could respond to different signals, integrate to the TFL1, AP1 and FT genes, and then synergistically regulate flowering in loquat.  Thus, this study provides a new insight into flowering regulation mechanisms in loquat.
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The effect of lactic acid bacteria inoculums on in vitro rumen fermentation, methane production, ruminal cellulolytic bacteria populations and cellulase activities of corn stover silage
GUO Gang, SHEN Chen, LIU Qiang, ZHANG Shuan-lin, SHAO Tao, WANG Cong, WANG Yong-xin, XU Qing-fang, HUO Wen-jie
2020, 19 (3): 838-847.   DOI: 10.1016/S2095-3119(19)62707-3
Abstract138)      PDF in ScienceDirect      
The objective of this study was to investigate the effect of lactic acid bacteria (LAB) inoculums on fermentation quality and in vitro digestibility of corn stover silage. Corn stover was ensiled without (control) or with Lactobacillus plantarum (LP), Enterococcus faecalis (EF), and Enterococcus mundtii (EM) for 45 days. The fermentation characteristics were assessed, and subsequent in vitro dry matter digestibility (DM-D), neutral detergent fiber digestibility (NDF-D), volatile fatty acids (VFA), methane (CH4 ) production, cellulolytic bacteria proportions and their activities per corn stover silage were also determined. There was no significant difference (P>0.05) among the silage pH, lactic acid, crude protein (CP), water soluble carbohydrates (WSC) and lignocelluloses contents of different treatments. The relative proportions of Ruminococcus flavefaciens and Fibrobacter succinogenes, carboxymethyl-ocellulose and β-glycosidase activities, DM-D, NDF-D, and VFA production of in vitro incubation was higher (P<0.05) for silages inoculated with LP and EF than those of the control silage. Silage inoculated with LP showed the lowest (P<0.05) CH4 production per unit yield of VFA, which was positively corresponded to the lowest (P<0.05) ratio of acetate to propionate. In summary, the ensiling fermentation quality and subsequent utilization of corn stover silage were efficiently improved by inoculated with L. plantarum.
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Yield-related agronomic traits evaluation for hybrid wheat and relations of ethylene and polyamines biosynthesis to filling at the mid-grain filling stage
YANG Wei-bing, QIN Zhi-lie, SUN Hui, LIAO Xiang-zheng, GAO Jian-gang, WANG Yong-bo, HOU Qi-ling, CHEN Xian-chao, TIAN Li-ping, ZHANG li-ping, MA Jin-xiu, CHEN Zhao-bo, ZHANG Feng-ting, ZHAO Chang-ping
2020, 19 (10): 2407-2418.   DOI: 10.1016/S2095-3119(19)62873-X
Abstract106)      PDF in ScienceDirect      
Because of the yield increase of 3.5–15% compared to conventional wheat, hybrid wheat is considered to be one of the main ways to greatly improve the wheat yield in the future.  In this study, we performed a principal component analysis (PCA) on two-line hybrids wheat and their parents using the grain weight (GW), the length of spike (LS), the kernel number of spike (KSN), and spike number (SPN) as variables.  The results showed that the variables could be classified into three main factors, the weight factor (factor 1), the quantity factor 1 (factor 2) and the quantity factor 2 (factor 3), which accounted for 37.1, 22.6 and 18.5%, respectively of the total variance in different agronomic variables, suggesting that the GW is an important indicator for evaluating hybrid combinations, and the grain weight of restorer line (RGW) could be used as a reference for parents selection.  The hybrid combination with a higher score in factor 1 direction and larger mid-parent heterosis (MPH) of the GW and its parents were used to carry out the analysis of grain filling, 1-aminocylopropane-1-carboxylicacid (ACC) and polyamine synthesis related genes.  The results suggested that the GW of superior grain was significantly higher than that of inferior grains in BS1453×JS1 (H) and its parents.  Both grain types showed a weight of H between BS1453 (M) and JS1(R), and a larger MPH, which may be caused by their differences in the active filling stage and the grain filling rate.  The ADP-glucose pyrophosphorylase (AGPase), granule bound starch synthase I (GBSSI), starch synthase III (SSS), and starch branching enzyme-I (SBE-I) expression levels of H were intermediated between M and R, which might be closely related to MPH formation of the GW.  Compared with R and H, the GW of M at maturity was the lowest.  The expression levels of spermidine synthase 2 (Spd2), ornithine decarboxylase (ODC) and S-adenosylmethionine decarboxylase (SAMDC) had significantly positive correlations with the grain filling rate (r=0.77*, 0.51*, 0.59*), suggesting their major roles in the grain filling and heterosis formation.  These provide a theoretical basis for improving the GW of photo-thermo-sensitive male sterile lines (PTSMSL) by changing the endogenous polyamine synthesis in commercial applications.
  
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Molecular cloning and functional identification of an apple flagellin receptor MdFLS2 gene
QI Chen-hui, ZHAO Xian-yan, JIANG Han, LIU Hai-tao, WANG Yong-xu, HU Da-gang, HAO Yu-jin
2018, 17 (12): 2694-2703.   DOI: 10.1016/S2095-3119(18)62009-X
Abstract266)      PDF (4711KB)(531)      
The leucine-rich repeat receptor kinase flagellin-sensing 2 gene (MdFLS2; Gene ID: MDP0000254112) was cloned from Royal Gala apple (Malus×domestica Borkh.).  This gene contained a complete open reading frame of 3 474 bp that encoded 1 158 amino acids.  The phylogenetic tree indicated that Prunus persica FLS2 exhibited the highest sequence similarity to MdFLS2.  The PlantCare database suggests that the promoter sequence of MdFLS2 contains several typical cis-acting elements, including ethylene-, gibberellin-, salicylic acid-, and drought-responsive elements.  Quantitative real-time PCR analysis showed that MdFLS2 was widely expressed in the different tissues of the apple and most highly expressed in the leaves.  Furthermore, MdFLS2 was significantly induced by the flagellin elicitor peptide flg22.  Treatment of the apple seedling leaves with flg22 resulted in an increase in leaf callose levels with increased treatment duration.  An increase in the production of O2 along with the expression of disease-related genes was also observed.  An oxidative burst was detected in the treated seedlings, but not in the control seedlings, indicating that flg22 had stimulated the expression of the MdFLS2 gene and its downstream target genes.  Furthermore, the ectopic expression of MdFLS2 complemented the function of the Arabidopsis fls2 mutant and conferred enhanced flg22 tolerance to the transgenic Arabidopsis, suggesting that MdFLS2 acts as a positive regulator in the response to pathogens in apple.
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Fine mapping of powdery mildew resistance gene PmTm4 in wheat using comparative genomics
XIE Jing-zhong, WANG Li-li, WANG Yong, ZHANG Huai-zhi, ZHOU Sheng-hui, WU Qiu-hong, CHEN Yong-xing, WANG Zhen-zhong, WANG Guo-xin, ZHANG De-yun, ZHANG Yan, HU Tie-zhu, LIU Zhi-yong
2017, 16 (03): 540-550.   DOI: 10.1016/S2095-3119(16)61377-1
Abstract1388)      PDF in ScienceDirect      
Powdery mildew, caused by Blumeria graminis f. sp. tritici, is one of the most severe wheat diseases.  Mining powdery mildew resistance genes in wheat cultivars and their appliance in breeding program is a promising way to control this disease.  Genetic analysis revealed that a single dominant resistance gene named PmTm4 originated from Chinese wheat line Tangmai 4 confers resistance to prevailing isolates of B. graminis f. sp. tritici isolate E09.  Detailed comparative genomics analyses helped to develop closely linked markers to PmTm4 and a fine genetic map was constructed using large F2 population, in which PmTm4 was located into a 0.66-cM genetic interval.  The orthologous subgenome region of PmTm4 in Aegilops tauschii was identified, and two resistance gene analogs (RGA) were characterized from the corresponding sequence scaffolds of Ae. tauschii draft assembly.  The closely linked markers and identified Ae. tauschii orthologs in the mapping interval provide an entry point for chromosome landing and map-based cloning of PmTm4.
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Identification of a major QTL for flag leaf glaucousness using a high-density SNP marker genetic map in hexaploid wheat
LI Chun-lian, LI Ting-ting, LIU Tian-xiang, SUN Zhong-pei, BAI Gui-hua, JIN Feng, WANG Yong, WANG Zhong-hua
2017, 16 (02): 445-453.   DOI: 10.1016/S2095-3119(16)61339-4
Abstract1030)      PDF in ScienceDirect      
Cuticular wax plays an important role in protecting land plant against biotic and abiotic stresses.  Cuticular wax production on plant surface is often visualized by a characteristic glaucous appearance.  This study identified quantitative trait loci (QTLs) for wheat (Triticum aestivum L.) flag leaf glaucousness (FLG) using a high-density genetic linkage map developed from a recombinant inbred line (RIL) population derived from the cross Heyne×Lakin by single-seed descent.  The map consisted of 2 068 single nucleotide polymorphism (SNP) markers and 157 simple sequence repeat (SSR) markers on all 21 wheat chromosomes and covered a genetic distance of 2 381.19 cM, with an average marker interval of 1.07 cM. Two additive QTLs for FLG were identified on chromosomes 3AL and 2DS with the increasing FLG allele contributed from Lakin.  The major QTL on 3AL, QFlg.hwwgr-3AL, explained 17.5–37.8% of the phenotypic variation in different environments.  QFlg.hwwgr-3AL was located in a 4.4-cM interval on chromosome 3AL that was flanked by two markers IWA1831 and IWA8374.  Another QTL for FLG on 2DS, designated as QFlg.hwwgr-2DS which was identified only in Yangling in 2014 (YL14), was flanked by IWA1939 and Xgwm261 and accounted for 11.3% of the phenotypic variation for FLG.  QFlg.hwwgr-3AL and QFlg.hwwgr-2DS showed Additive×Environment (AE) interactions, explaining 3.5 and 4.4% of the phenotypic variance, respectively.  Our results indicated that different genes/QTLs may contribute different scores of FLG in a cultivar and that the environment may play a role in FLG.
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The variation of NAD+-SDH gene in mutant white-fleshed loquat
LI Jing, WANG Yong-qing, CHEN Dong, TU Mei-yan, XIE Hong-jiang, JIANG Guo-liang, LIU Jia, SUN Shu-xia
2016, 15 (8): 1744-1750.   DOI: 10.1016/S2095-3119(15)61297-7
Abstract1833)      PDF in ScienceDirect      
   Loquat (Eriobotrya japonica Lindl.) can be divided into yellow- and white-fleshed cultivars by flesh color. However, a Dongting loquat mutant, which involved bud sport and growing white-fleshed fruit in the central region of the trunk (as wild loquat bears yellow-fleshed fruits naturally), was discovered in the preliminary study. The study cloned the coding sequence (CDS) of NAD+-dependent sorbitol dehydrogenase (NAD+-SDH ) gene from the selected materials of mutant loquat, wild loquat and other nine loquat cultivars/accessions, and found that the CDS of NAD+-SDH gene from the mutant loquat, other than the rest two types of materials, had three single nucleotide polymorphisms (SNPs) loci; in addition, the amino acid encoded at variation loci changed accordingly. NAD+-SDH plays an active role in converting sorbitol into fructose in loquat cultivars. For the mutant white-fleshed loquat, the activity of NAD+-SDH rises first and then drops, the sorbitol content decreases steadily, and its fructose content is higher than that in wild loquat from coloration to maturation stage. As demonstrated by the real-time fluorescence quantification PCR analysis, the expression level of NAD+-SDH gene at maturation stage is about 5-fold lower than wild type. It may be assumed that, the three SNPs loci might lead to excessive conversion of sorbitol into fructose under the catalytic action of NAD+-SDH of white-fleshed mutant loquat at maturation stage, resulting in the increase of fructose content and reduced expression abundance of mRNA after transcription. Besides, NAD+-SDH gene may be related to flesh color and carbohydrate variation of white-fleshed mutant loquat.
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Effects of graded fiber level and caecectomy on metabolizable energy value and amino acid digestibility in geese
YANG Jing, ZHAI Shuang-shuang, WANG Yong-chang, WANG Shen-shen, YANG Zhi-peng, YANG Lin
2016, 15 (3): 629-635.   DOI: 10.1016/S2095-3119(15)61071-1
Abstract1994)      PDF in ScienceDirect      
This study was carried out to evaluate the effects of graded levels of fiber from rice hull and the caecectomy on metabolizable energy (ME) and amino acid digestibility (AAD) in 42 Wuzong geese (21 intact and 21 caecectomized). The experiment was a 3×2 factorial randomized design with 3 fiber levels and 2 treatments of geese (caecectomized and intact). Each of the 3 diets was tube-fed (60 g) to 7 caecectomized and 7 intact adult Wuzong geese that had been fasted for 24 h. Excreta were collected for the next 48 h. The same geese were also fasted for estimation of endogenous losses of amino acids (AA) and energy after short period recovery. AA content and metabolizable energy value were determined in the excreta. In general, significant effects were found for fiber levels on apparent digestibility of Thr, Leu, Pro, Glu,Val, Ala and total AA (P<0.05). Apparent metabolizable energy (AME) and true metabolizable energy (TME) were both significantly influenced by increased fiber levels (P<0.01). Compared with caecectomized geese, markedly increases of AME and TME were found in intact geese (P<0.01 for AME and P<0.05 for TME), but no difference in the AAD were found between intact and caecectomized geese (P>0.05), except for Lys and Try (P<0.05). These observations suggested that dietary fiber level played an important role in digestibility of AAD and ME. Intact geese should be chosen for the nutrient evaluation of feedstuffs in practice, resulting from the significant difference between caecectomized and intact geese.
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Comparative genetic mapping revealed powdery mildew resistance gene MlWE4 derived from wild emmer is located in same genomic region of Pm36 and Ml3D232 on chromosome 5BL
ZHANG Dong, OUYANG Shu-hong, WANG Li-li, CUI Yu, WU Qiu-hong, LIANG Yong, WANG Zhen-zhong, XIE Jing-zhong, ZHANG De-yun, WANG Yong, CHEN Yong-xing, LIU Zhi-yong
2015, 14 (4): 603-609.   DOI: 10.1016/S2095-3119(14)60774-7
Abstract1674)      PDF in ScienceDirect      
Powdery mildew, caused by Blumeria graminis f. sp. tritici, is one of the most devastating wheat diseases. Wild emmer wheat (Triticum turgidum ssp. dicoccoides) is a promising source of disease resistance for wheat. A powdery mildew resistance gene conferring resistance to B. graminis f. sp. tritici isolate E09, originating from wild emmer wheat, has been transferred into the hexaploid wheat line WE4 through crossing and backcrossing. Genetic analyses indicated that the powdery mildew resistance was controlled by a single dominant gene, temporarily designated MlWE4. By mean of comparative genomics and bulked segregant analysis, a genetic linkage map of MlWE4 was constructed, and MlWE4 was mapped on the distal region of chromosome arm 5BL. Comparative genetic linkage maps showed that genes MlWE4, Pm36 and Ml3D232 were co-segregated with markers XBD37670 and XBD37680, indicating they are likely the same gene or alleles in the same locus. The co-segregated markers provide a starting point for chromosome landing and map-based cloning of MlWE4, Pm36 and Ml3D232.
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Analysis of the function of D279N mutation of VP2 of infectious bursal disease virus
QI Xiao-le, LU Zhen, WANG Nian, CHEN Yu-ming, ZHANG Li-zhou, GAO Li, LI Kai, REN Xian-gang, WANG Yong-qiang, GAO Hong-lei, GAO Yu-long, Nicolas Eterradossi, WANG Xiao-mei
2015, 14 (12): 2618-2625.   DOI: 10.1016/S2095-3119(14)60933-3
Abstract1405)      PDF in ScienceDirect      
Infectious bursal disease virus (IBDV) is responsible for the highly contagious infectious bursal disease of chickens. Further understanding the gene-function is necessary to design the tailored vaccine. The amino acid residue 279, located on strand PF of VP2, is one of the three residues that have been reported to be involved in cell-tropism but with some inconsistency. In this study, to further clarify the amino acids involved in the cell tropism of IBDV, a series of mutations about residue 279 were introduced into the VP2 of vvIBDV Gx strain. With the reverse genetic system, we found single mutation of D279N, double mutations of D279N/A284T or Q253H/D279N were not enough to adapt IBDV to chicken embryo fibroblast (CEF) cell. To evaluate whether residue 279 could influence the replication and virulence of IBDV, the virus rGxHT-279 with three mutations (Q253H/D279N/A284T) was rescued and evaluated. Results showed that the mutation of residue 279 in VP2 had no efficient effects on both the replication efficiency in vitro and the virulence to SPF chickens of IBDV. In summary, the results demonstrated that residue 279 of VP2 did not contribute efficiently to cell tropism, replication efficiency, and virulence of IBDV at least in some strains. These findings provided further information for understanding the gene function of IBDV.
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A Dominant Locus, qBSC-1, Controls β Subunit Content of Seed Storage Protein in Soybean (Glycine max (L.) Merri.)
WANG Jun, LIU Lin, GUO Yong, WANG Yong-hui, ZHANG Le, JIN Long-guo, GUAN Rong-xia, LIU Zhang-xiong, WANG Lin-lin, CHANG Ru-zhen , QIU Li-juan
2014, 13 (9): 1854-1864.   DOI: 10.1016/S2095-3119(13)60579-1
Abstract1422)      PDF in ScienceDirect      
Soybean seed storage protein is one of the most important plant vegetable proteins, and β subunit is of great significance to enhance soybean protein quality and processing property. F2 segregated population and residual heterozygous lines (RHL) derived from the cross between Yangyandou (low level of β subunit) and Zhonghuang 13 (normal level of β subunit) were used for mapping of β subunit content. Our results showed that β subunit content was controlled by a single dominant locus, qBSC-1 (β subunit content), which was mapped to a region of 11.9 cM on chromosome 20 in F2 population of 85 individuals. This region was narrowed down to 2.5 cM between BARCSOYSSR_20_0997 and BARCSOYSSR_20_0910 in RHL with a larger population size of 246 individuals. There were 48 predicted genes within qBSC-1 region based on the reference genome (Glyma 1.0, Williams 82), including the two copies of β subunit coding gene CG4. An InDel marker developed from a thymine (TT) insertion in one copy of CG4 promoter region in Yangyandou cosegregrated with BARCSOYSSR_20_0975 within qBSC-1 region, suggesting that this InDel marker maybe useful for marker-assisted selection (MAS).
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17β-Estradiol Regulates SKP2 Expression in Cultured Immature Boar Sertoli Cells Mainly via Estrogen Receptor β, cAMP-PKA and ERK1/2
WANG Xian-zhong, ZHU Feng-wei, WANG Yong, WANG Yi, ZHANG Jiao-jiao , ZHANG Jia-hua
2014, 13 (4): 827-836.   DOI: 10.1016/S2095-3119(13)60430-X
Abstract1748)      PDF in ScienceDirect      
Estrogen plays an important role in regulating testicular Sertoli cell number. Furthermore, S-phase kinase-associated protein 2 (SKP2) plays a central role in mammalian cell cycle progression. The objective of this study was to determine whether 17β-estradiol can regulate the expression of SKP2, and the Sertoli cell cycle, via estrogen receptor β (ERβ), the cyclic adenosine monophosphate (cAMP)-protein kinase A (PKA) and extracellular signal-regulated kinase (ERK1/2) pathway. When cultured immature boar Sertoli cells were treated with 17β-estradiol, a time-dependent increase in SKP2 mRNA and protein level was observed by real-time PCR and Western blot, and 17β-estradiol activity peaked at 30 min. Treatment with ICI182780 and ERβ antagonist reduced 17β-estradiol-induced expression of SKP2 and proliferating cell nuclear antigen (PCNA), while increasing the protein concentration of p27kip1. However, the effect of ERa antagonist on these parameters was lower than that of ICI182780 and ERβ. Forskolin had a similar effect as 17β-estradiol on the expression of SKP2, PCNA and p27kip1. Rp-cAMP, H-89 and U0126 treatment reduced 17β-estradiol-induced changes, while H-89 also inhibited ERK1/2 activation. Therefore, 17β-estradiol mainly regulates SKP2 mRNA and protein expression via ERβ-cAMP-PKA and ERK1/2 activation. SKP2 and PCNA expression were positively correlated, while increased SKP2 expression likely resulted in p27kip1 degradation.
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Effects of High-Quality Aromatic Rice Varieties on the Fitness of the Striped Stem Borer, Chilo suppressalis (Walker) in Central China
DU Xue-zhu, WANG Yong, CHEN Long-jia, PENG Chuan-hua, MA Wei-hua, LEI Chao-liang
2013, 12 (7): 1208-1214.   DOI: 10.1016/S2095-3119(13)60446-3
Abstract1944)      PDF in ScienceDirect      
The striped stem borer (SSB), Chilo suppressalis (Walker) (Lepidoptera: Pyralidae), is a major pest of rice in China. Variation in host-plant quality may affect the body size of herbivorous insects, which in turn, can determine their lifehistory parameters such as survival, fecundity and fitness. In this study, we tested the effects of high-quality aromatic rice varieties on the fitness of SSB in China. Results showed that 1st instar larvae had higher penetrating rates and survival rates on the high-quality aromatic rice varieties (EX-1 and WX-988) than that on the non-aromatic rice varieties (EZ-5 and LYP-9). In addition, shorter developmental periods, greater female pupal weights and higher of other life-history parameters (hatching rate, pupation rate, eclosion rate and increase index) were found on the high-quality aromatic rice varieties, although only female pupal weight showed a significant difference between the two varieties. The highest dead heart rate was found on the aromatic rice variety of EX-1. These results indicate that SSB sustains a lower fitness cost when consuming the high-quality aromatic rice varieties (EX-1 and WX-988) than on the non-aromatic rice varieties (EZ-5 and LYP-9) in Central China.
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Assessment of Genetic Relationship of Foxtail Millet with Its Wild Ancestor and Close Relatives by ISSR Markers
LI Wei, ZHI Hui, WANG Yong-fang, LI Hai-quan , DIAO Xian-min
2012, 12 (4): 556-566.   DOI: 10.1016/S1671-2927(00)8575
Abstract1710)      PDF in ScienceDirect      
Inter simple sequence repeat (ISSR) analysis was applied to samples of foxtail millet and its wild ancestor and other close relatives of the genus Setaria in order to detect domestication-related geographical structure and phylogenetic relationship of those species. Eighty-one accessions of nine Setaria species that originated from different regions were used in this study. Fourteen out of the 27 ISSR primers screened amplified successfully and a total of 156 markers were scored for all the accessions, with a high level of polymorphism being detected. The dendrogram based on UPGMA cluster analysis showed clear geographic structure among foxtail millet and its wild ancestor green foxtail, which implies that northern China is the domestication center for both the Chinese and European foxtail millet landraces used in this study. This result did not support the hypothesis that China and Europe are independent domestication centers for foxtail millet proposed by several previous studies based on morphological and isozyme data. The dendrogram also clearly classified the Setaria sample used into two groups, a viridis and a pumila groups. The viridis group was composed of S. viridis, S. italica, S. faberii, S. verticillata, S. leucopila, and S. queenslandica, and the pumila group consisted of S. parviflora and S. pumila, which is consistent with the recently proposed hypothesis of multiple origin of Setaria species. The phylogenetic relationships among different species are discussed.
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Effects of the Residues of Cuscuta campestris and Mikania micrantha on Subsequent Plant Germination and Early Growth
LI Feng-lan, LI Ming-guang, ZAN Qi-jie, GUO Qiang, ZHANG Wei-yin, WU Zhi, WANG Yong-jun
2012, 12 (11): 1852-1860.   DOI: 10.1016/S1671-2927(00)8720
Abstract1574)      PDF in ScienceDirect      
Cuscuta campestris, a dodder, can parasitize and suppress a scrambling herbaceous to semi-woody perennial vine, Mikania micrantha, one of the most destructive weeds in the world. To assess the effects of the mixed residue of C. campestris and M. micrantha on the subsequent plant community, we conducted a one-year experiment on the germination and seedling growth of subsequent plant community after the application of C. campestris. Seven treatments of varying proportions of C. campestris and M. micrantha residue on 21 subject trees and shrubs, which were commonly found in South China, resulted in a germination rate of 35.3% for all 8715 seeds from 18 species, ranging from 5.7 to 81.9%; the remaining 3 species failed to germinate. ANOVA analysis showed that the residue did not affect the germination, growth, or mortality of the trees and shrubs. The germinated C. campestris seeds from the residue coiled the seedlings of most of the species, but less than 4% host death caused by C. campestris. In addition, the residue did not affect the germination of the herbaceous seedlings originating from the loam, and the similarity coefficients of the germinated seedlings between the treatments were very high. These results suggested that the residue had no negative impact on the germination and early seedling stages of the tree, shrub and grass species of the subsequent plant community. The use of C. campestris residue had a positive effect on the growth of M. micrantha, but it did not change the trend of M. micrantha being suppressed because re-parasitization occurred soon after the growth restarted. No negative effect was detected on the other species as a result of the parasitization of C. campestris or by the use of the mixed residue. This suggests that C. campestris is likely to be an effective and promising ecologically safe native herbaceous agent for controlling M. micrantha.
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Effects of Extreme Air Temperature and Humidity on the Insecticidal Expression Level of Bt Cotton
CHEN Yuan, WEN Yu-jin, CHENYuan , John Tom Cothren, ZHANG Xiang, WANG Yong-hui, William A
2012, 12 (11): 1836-1844.   DOI: 10.1016/S1671-2927(00)8718
Abstract1265)      PDF in ScienceDirect      
The higher survival rates of Helicoverpa amigera larvae were usually observed after adverse climate which was related to extreme temperature (T) and relative humidity (RH) stresses in transgenic Bacillus thuringiensis (Bt) cotton. The unstable resistance of Bt cotton to bollworms has been correlated with the reduced expression of CryIAc d-endotoxin. The objective of this study was to investigate the effects of combined temperature and relative humidity stresses on the leaf CryIAc insecticidal protein expression during critical developmental stages. The study was undertaken on two transgenic cotton cultivars that share same parental background, Sikang 1 (a conventional cultivar) and Sikang 3 (a hybrid cultivar), during the 2007 and 2008 growing seasons at the Yangzhou University Farm, Yangzhou, China. The study was arranged with two factors that consisted of temperature (two levels) and relative humidity (three levels). The six T/RH treatments were 37°C/95%, 37°C/70%, 37°C/50%, 18°C/95%, 18°C/70%, and 18°C/50%. In 2007, the six treatments were imposed to the plants at peak flowering stage for 24 h; in 2008, the six treatments were applied to the plants at peak square, peak flowering, and peak boll stages for 48 h. The results of the study indicated that the leaf insecticidal protein expression in CryIAc was significantly affected by extreme temperature only at peak flowering stage, and by both extreme temperature and relative humidity during boll filling stage. The greatest reductions were observed when the stresses were applied at peak boll stage. In 2008, after 48 h stress treatment, the leaf Bt endotoxin expression reduced by 25.9-36.7 and 23.6-40.5% at peak boll stage, but only by 14.9-26.5 and 12.8-24.0% at peak flowering stage for Sikang 1 and Sikang 3, respectively. The greatest reduction was found under the low temperature combined with low relative humidity condition for both years. It is believed that the temperature and relative humidity stresses may be attributed to the reduced efficacy of Bt cotton in growing conditions in China, where extreme temperatures often increase up to 35-40°C and/or decrease down to 15-20°C, and relative humidity may reach to 85-95% and/or reduce to 40-55% during the cotton growing season.
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