Field effect of Cnaphalocrocis medinalis granulovirus (CnmeGV) on the pest of rice leaffolder

Rice leaffolder, Cnaphalocrocis medinalis(Guenée), has become a major pest throughout the rice cultivating areas of China and caused severe damage to rice production. Cnaphalocrocis medinalis granulovirus(CnmeGV), a naturally occurring baculovirus, is revealed as a potential microbial agent for the pest control. Field applications of CnmeGV were conducted against rice leaffolder larvae in rice paddies. CnmeGV infected the larvae not only in the current generation but also in the successive generation, resulting in a sustained infection in the larva population for at least 48 days. Under diferent concentrations of CnmeGV(7.5×10~(11) and 1.125×10~(12) occlusion body(OB) ha~(-1)) at 30 days after spraying, larval population reduced up to 76.32% and rice leaf rolled rate kept in 15.42%. Simultaneously, CnmeGV had no impact on arthropod predators of C. medinalis, with abundances ranging from 2.39 to 3.79 per ten hills. These results revealed that CnmeGV is suitable as a bio-pesticide for rice leaffolder management in rice paddies.     

Decreased panicle N application alleviates the negative effects of shading on rice grain yield and grain quality

Light deficiency is a growing abiotic stress in rice production. However, few studies focus on shading effects on grain yield and quality of rice in East China. It is also essential to investigate proper nitrogen (N) application strategies that can effectively alleviate the negative impacts of light deficiency on grain yield and quality in rice. A two-year field experiment was conducted to explore the effects of shading (non-shading and shading from heading to maturity) and panicle N application (N_DP, decreased panicle N rate; N_MP, medium panicle N rate; N_IP, increased panicle N rate) treatments on rice yield- and quality-related characteristics. Compared with non-shading, shading resulted in a 9.5–14.8% yield loss (P<0.05), mainly due to lower filled-grain percentage and grain weight. N_MP and N_IP had higher (P<0.05) grain yield than N_DP under non-shading, and no significant difference was observed in rice grain yield among N_DP, N_MP, and N_IP under shading. Compared with N_MP and N_IP, N_DP achieved less yield loss under shading because of the increased filled-grain percentage and grain weight. Shading reduced leaf photosynthetic rate after heading, as well as shoot biomass weight at maturity, shoot biomass accumulation from heading to maturity, and nonstructural carbohydrate (NSC) content in the stem at maturity (P<0.05). The harvest index and NSC remobilization reserve of N_DP were increased under shading. Shading decreased (P<0.05) percentages of brown rice, milled rice, head rice, and amylose content while increasing (P<0.05) chalky rice percentage, chalky area, chalky degree, and grain protein. N_MP demonstrated a better milling quality under non-shading, while N_DP demonstrated under shading. N_DP exhibited both lower chalky rice percentage, chalky area, and chalky degree under non-shading and shading, compared with N_MP and N_IP. N_DP under shading decreased amylose content and breakdown but increased grain protein content and setback, contributing to similar overall palatability to non-shading. Our results suggested severe grain yield and quality penalty of rice when subjected to shading after heading. N_DP improved NSC remobilization, harvest index, and sink-filling efficiency and alleviated yield loss under shading. Besides, N_DP would maintain rice’s milling, appearance, and cooking and eating qualities under shading. Proper N management with a decreased panicle N rate could be adopted to mitigate the negative effects of shading on rice grain yield and quality.     

Mapping and genetic validation of a grain size QTL qGS7.1 in rice(Oryza sativa L.)

Grain size is a major determinant of grain weight, which is one of the components of rice yield. The objective o this study was to identify novel, and important quantitative trait loci(QTLs) for grain size and weight in rice. QTLs were mapped using a BC_4F_4 population including 192 backcross inbred lines(BILs) derived from a backcross between Xiaolijing(XLJ) and recombinant inbred lines(RILs). The mapping population was planted in both Lingshui(Hainan, 2015) and Fuyang(Zhejiang, 2016), with the short-and long-day conditions, respectively. A total of 10 QTLs for grain length, four for grain width, four for the ratio of grain length to width, and 11 for grain weight were detected in at least one environment and were distributed across 11 chromosomes. The phenotypic variance explained ranged from 6.76–25.68%, 14.30–34.03%, 5.28–26.50%, and 3.01–22.87% for grain length, grain width, the ratio of grain length to width, and thousand grain weight, respectively. Using the sequential residual heterozygotes(SeqRHs) method, qGS7.1, a QTL for grain size and weight, was mapped in a 3.2-Mb interval on chromosome 7. No QTLs about grain size and weight were reported in previous studies in this region, providing a good candidate for functional analysis and breeding utilization.     

Reduction in cadmium accumulation in japonica rice grains by CRISPR/Cas9-mediated editing of OsNRAMP5

Cadmium (Cd) intake is harmful to human health and Cd contamination in rice grains represents a severe threat to those consuming rice as a staple food. Knockout of Cd transporters is a promising strategy to reduce Cd accumulation in rice grains. OsNRAMP5 is the major transporter for Cd and manganese (Mn) uptake in rice. Nevertheless, it is uncertain whether knockout of OsNRAMP5 is applicable to produce low Cd rice without affecting plant growth and grain yield. In this study, we adopted CRISPR/Cas9-based gene editing technology to knock out OsNRAMP5 in two japonica varieties. We generated three independent transgene-free osnramp5 mutants and investigated the effect of osnramp5 mutations on Cd accumulation and plant growth. Hydroponic experiments showed that plant growth and chlorophyll content were significantly reduced in osnramp5 mutants at low Mn conditions, and this defective growth in the mutants could be fully rescued by supply of high levels of Mn. Cd and Mn accumulation in both roots and shoots was markedly reduced in the mutants compared to that in wild-type plants. In paddy field experiments, although Cd in flag leaves and grains was greatly reduced in osnramp5 mutants, some agronomic traits including plant height, seed setting rate, and grain number per panicle were affected in the mutants, which ultimately caused a mild reduction in grain yield. The reduced plant growth in the mutants can be attributed to a marked decrease in Mn accumulation. Our results reveal that the manipulation of OsNRAMP5 should be treated with caution: When assessing the applicability of osnramp5 mutants, soil pH and soil water content in paddy fields need to be taken into consideration, since they might affect the levels of available Mn in the soil and consequently determine the effect of the mutation on grain yield.     

Estimates on nitrogen uptake in the subsequent wheat by above-ground and root residue and rhizodeposition of using peanut labeled with 15N isotope on the North China Plain

Leguminous crops play a vital role in enhancing crop yield and improving soil fertility. Therefore, it can be used as an organic N source for improving soil fertility. The purpose of this study was to (i) quantify the amounts of N derived from rhizodeposition, root and above-ground biomass of peanut residue in comparison with wheat and (ii) estimate the effect of the residual N on the wheat-growing season in the subsequent year. The plants of peanut and wheat were stem fed with ¹⁵N urea using the cotton-wick method at the Wuqiao Station of China Agricultural University in 2014. The experiment consisted of four residue-returning strategies in a randomized complete-block design: (i) no return of crop residue (CR0); (ii) return of above-ground biomass of peanut crop (CR1); (iii) return of peanut root biomass (CR2); and (iv) return of all residue of the whole peanut plant (CR3). The 31.5 and 21% of the labeled ¹⁵N isotope were accumulated in the above-ground tissues (leaves and stems) of peanuts and wheat, respectively. N rhizodeposition of peanuts and wheat accounted for 14.91 and 3.61% of the BG¹⁵N, respectively. The ¹⁵N from the below-ground ¹⁵N-labeled of peanuts were supplied 11.3, 5.9, 13.5, and 6.1% of in the CR0, CR1, CR2, and CR3 treatments, respectively. Peanut straw contributes a significant proportion of N to the soil through the decomposition of plant residues and N rhizodeposition. With the current production level on the NCP, it is estimated that peanut straw can potentially replace 104 500 tons of synthetic N fertilizer per year. The inclusion of peanut in rotation with cereal can significantly reduce the use of N fertilizer and enhance the system sustainability.     

Residue management induced changes in soil organic carbon and total nitrogen under different tillage practices in the North China Plain

Crop residue retention has been considered a practicable strategy to improve soil organic carbon(SOC) and total nitrogen(TN), but the effectiveness of residue retention might be different under varied tillage practices. To evaluate the effects of residue management on the distribution and stocks of SOC and TN under different tillage practices, a bifactorial experiment with three levels for tillage practices(no-tillage, rotary tillage, and conventional tillage) and two levels for residue managements(residue retention and residue removal) was conducted in the North China Plain(NCP). Results showed that after a short experimental duration(3–4 years), concentrations of SOC and TN in the 0–10 cm layer were higher under no-tillage than under conventional tillage, no matter whether crop residues were retained or not. Residue retention increased SOC and TN concentrations in the upper layers of soil to some degree for all tillage practices, as compared with residue removal, with the greatest increment of SOC concentration occurred in the 0–10 cm layer under rotary tillage, but in the 10–30 cm layer under conventional tillage. The stocks of SOC in the 0–50 cm depth increased from 49.89 Mg ha–1 with residue removal to 53.03 Mg ha–1 with residue retention. However, no-tillage did not increase SOC stock to a depth of 50 cm relative to conventional tillage, and increased only by 5.35% as compared with rotary tillage. Thus, residue retention may contribute more towards SOC sequestration than no-tillage. Furthermore, the combination between residue retention and no-tillage has the greatest advantage in enhancing SOC and TN in the NCP region.     

不同播期与收获期对水稻灌浆期、产量及米质的影响

【目的】明确不同水稻品种(系)的适宜播种时间,确保水稻生产的优质高产。【方法】以6个恢复系及其所配的12个组合为材料,采用4期播种(前后两期间隔6 d),利用二因素两向分组资料无重复观察值模型与最小显著差数测验法(LSD法)方差分析和显著性比较,研究不同播期对籽粒产量、灌浆期、加工品质和外观品质的影响,以及成熟后推迟收获对碾米品质和外观品质的影响。【结果】不同播期的恢复系糙米率有显著差异,随着播期的推迟糙米率有降低趋势;籽粒产量、灌浆期、精米率、整精米率、垩白率和垩白度在不同播期间无显著差异。II优组合不同播期间籽粒产量差异不显著,宜香优组合1~3期间籽粒产量差异不显著,第4期显著减少;不同播期对杂交稻的碾米品质和外观品质影响较大,推迟播种会降低糙米率、垩白率和垩白度,但对精米率和整精米率影响不显著。第4期推迟收获会导致糙米率、垩白率和垩白度增加,精米率和整精米率不同程度降低。【结论】恢复系和杂交稻在川南3月27日后推迟播种,产量有减产趋势,4月上中旬播种产量减少极显著;恢复系和杂交稻谷粒长宽比不受播期影响;杂交稻灌浆期、碾米品质和外观品质受播期的影响比恢复系大;杂交稻成熟后不及时收获会导致稻米加工品质和外观品质变差。     

Effects of the severity and timing of basal leaf removal on the amino acids profiles of Sauvignon Blanc grapes and wines

The effects of the severity and timing of leaf removal(LR) on the amino acids of Sauvignon Blanc grapes and wines were studied during the 2017 growing season. High-performance liquid chromatography(HPLC) was used to analyze the amino acids profiles of grape berries and wines. The basal leaves were removed at three time points(40, 56 and 72 days after flowering, named LR40, LR56 and LR72, respectively) at two severity levels(one at which the first, third, and fifth basal leaves of each shoot were removed(50% level); and another at which the first six basal leaves were removed(100% level)). The results showed that leaf removal had little impact on total soluble solids(°Brix), titratable acidity, pH or berry weight. The LR72-50% treated grapes had higher berry weight, titratable acidity and °Brix than those of the other treatments. The highest concentrations of total amino acids and of total amino acids except proline were detected in LR72-50% treated grapes(2 952.58 and 2 764.36 mg L~(-1), respectively); the lowest were detected in LR72-100% treated grapes(2 172.82 and 2 038.71 mg L~(-1), respectively). LR72-50% treatment significantly promoted the synthesis of aspartic acid, serine, arginine, alanine, aminobutyric acid and proline at both severity levels for grapes, the concentrations of all of these amino acids were increased relative to the control concentrations. The LR72-50%, LR40-100% and LR72-100% treated wines had higher total amino acids concentrations and higher concentrations of some individual amino acids, such as arginine, alanine and serine, than did the control wines. Of all the amino acids studied, glycine, tyrosine, cysteine, methionine and lysine were not significantly influenced by the timing or severity basal defoliation in grapes and wines. The present study reveals the effects of the timing and severity of leaf removal on the amino acids profiles of grapes and wines.     

Field identification of morphological and physiological traits in two special mutants with strong tolerance and high sensitivity to drought stress in upland rice(Oryza sativa L.)

The two mutants idr1-1 and 297-28, which were obtained from the radiation mutation of HD297 and IAPAR9, were used as experimental materials in this study for a 2-year(2012 and 2013) experiment about field drought resistance identification in Beijing, China. Key agronomic traits and water-related physiological indexes were observed and measured, including the leaf anti-dead level(LADL), days to heading, plant height, setting percentage, aboveground biomass, leaf water potential(LWP), net photosynthetic rate(Pn) and transpiration rate. The results showed that the mutant idr1-1 that was under drought stress(DS) conditions for 2 years had the highest LADL grades(1.3 and 2.0) among all the materials, and they were 2–3 grades stronger than the wild-type IAPAR9 with an average that was 21.4% higher for the setting percentage than the wild type. Compared with the IAPAR9 for the 2-year average delay in the days to heading and the reduction rates in the plant height, setting percentage, and aboveground biomass under DS compared with the well-watered(WW) treatment, idr1-1 showed 3.2% less delay and 19.1, 16.4, and 6.1% less reduction, respectively. The idr1-1 in the LWP always exhibited the highest performance among all the materials. The Pn of idr1-1 under severe and mild DS comparing with that under WW was slightly decreased and even slightly increased, respectively, leading to an average reduction rate of only 0.92%, which was 26.93% less than that of IAPAR9. Under the severe DS, idr1-1 still showed the highest value of 16.88 μmol CO2 m–2 s–1 among all the materials and was significantly higher than that of IAPAR9(11.66 μmol CO2 m–2 s–1). Furthermore, only idr1-1 had the increased and the highest transpiration rate values(7.6 and 6.04 mmol H2 O m–2 s–1) under both mild and severe DS compared with the values under WW, when the transpiration rate of all the other materials significantly decreased. By contrast, the 297-28 in terms of the LADL grade under DS was the lowest(7.0), and it was four grades weaker than its wildtype HD297 and even one grade weaker than the drought-sensitive paddy rice SN265. For the 2-year average reduction rates in aboveground biomass and plant heights under DS compared with those under the WW, 297-28 was 31.6 and 31.8% higher than HD297, respectively. Meanwhile, 297-28 showed the worst performance for the LWP, Pn, and transpiration rate. These results suggest that idr1-1 might be a superior drought tolerant mutant of upland rice found in China. It has a strong ability to maintain and even enhance leaf transpiration while maintaining a high plant water potential under DS, thus supporting a high Pn and alleviating the delay in agronomic trait development and yield loss effectively. 297-28 is a much more highly drought-sensitive mutant that is even more sensitive than paddy rice varieties. The two mutants could be used as drought tolerance controls for rice germplasm identification and the drought resistant mechanism studies in the future. idr1-1 is also suitable for breeding drought-tolerant and lodging-resistant high-yield rice varieties.     

灌浆结实阶段水分逆境对超级稻叶片丙二醛含量的影响

通过水份仪结合称重法人为控制土壤水分的盆栽试验,研究灌浆结实阶段水分逆境对不同类型超级稻品种叶片丙二醛(MDA)含量的影响.结果表明:水稻灌浆结实阶段对水分的敏感性较强,开花至花后30 d不同时段的水分逆境均导致叶片丙二醛含量显著增加,尤其花后30 d时叶片丙二醛含量较对照增加更为明显.不同水分条件下各品种倒二叶的丙二醛含量也均显著高于剑叶,水分逆境导致水稻叶片尤其是倒二叶的丙二醛含量显著升高,叶片膜脂过氧化程度增强,加速了叶片的衰老进程.不同品种由于对水分逆境响应的敏感性差异而受影响的程度不同,相对于普通杂交稻汕优63来说,超级稻品种受到的影响要小.     

Species-specific COI primers for rapid identification of a globally significant invasive pest,the cassava mealybug Phenacoccus manihoti Matile-Ferrero

The globally invasive cassava mealybug Phenacoccus manihoti Matile-Ferrero is a pernicious pest of cassava,and its recent introduction into Asia has raised considerable alarm.To slow or prevent further invasion,an accurate,simple,and developmental-stage-independent detection method for P.manihoti is required.In the present study,a PCR method based on a species-specific mitochondrial DNA cytochrome oxidase I(SS-COI)marker was developed for rapid identification of P.manihoti.One pair of SS-COI primers(PMSSZW-1F and PMSSZW-1R)was designed based on sequence variations in the COI gene among P.manihoti and related mealybug species.Specificity of the primer pair was validated on 21 closely related species.Sensitivity tests were performed on four immature developmental stages and female adults.Efficacy tests demonstrated that at the relatively low concentration of(135.2±14.7)pgresuspended DNA,the specific fragment was detected in all replicates.Furthermore,the SS-COI primer pair was assayed on three populations of P.manihoti from major exporting countries of cassava.The PCR assay was proved to be a rapid,simple,and reliable molecular measure for the identification of P.manihoti.This tool will be useful for quarantine,monitoring,and management of this invasive pest.     

A review: The beneficial effects and possible mechanisms of aluminum on plant growth in acidic soil

Aluminum(Al) is the third most abundant element in the earth's crust and a major factor inhibiting plant growth and reducing crop yield in acidic soil. Although there is substantial research on the phytotoxic effects and the underlying mechanisms of Al by applying Al alone hydroponically, soil is a complex medium containing numerous mineral elements that can interact with Al and other elements and their bioavailability in plants. In this review, we describe the roles of Al in promoting plant growth, enhancing phosphorus availability and efficient use in plants, and alleviating H+, iron, and manganese toxicity in acidic conditions. Furthermore, we discuss the possible mechanisms of enhanced abiotic stress tolerance induced by Al. We also elucidate the role of Al in attracting plant growth promoting rhizo-bacteria(PGPR) and their interactions with plants by increasing organic exudates.     

粳稻品质与株型的关系研究

1999-2000年在田间条件下，对粳稻品种（系）的叶片和茎秆形态、穗型及群体光合速率与稻米碾磨品质和外观品质的关系进行了研究。结果表明，顶部叶片、穗下节间、叶鞘长度、穗的着粒密度、群体光合速率与稻米品质各指标关系密切。理想的优质粳稻株型应为：在一定范围内，上部叶片长、窄，穗下节间长，中下部节间短、叶鞘长，着粒密度较小的散穗型，经济产量和品质形成期较弱的群体光合速率。     

Effect of exogenous GA_3 on flowering quality,endogenous hormones,and hormone-and flowering-associated gene expression in forcing-cultured tree peony(Paeonia suffruticosa)

Gibberellins(GAs) promote flowering in the forcing-cultured tree peony(Paeonia suffruticosa), however, the mechanism of regulating flowering is not fully understood. In this study, exogenous GA_3 was applied to five-year-old Luoyang Hong plants to explore responses in terms of endogenous hormones, flowering quality, and the hormone-and flowering-associated gene expression. Exogenous GA_3 application significantly promoted flower bud development and new branch growth, as well as improved flowering quality. Exogenous GA_3 application also stimulated the synthesis of endogenous GA_3 and indole-3-acetic acid(IAA) but reduced abscisic acid(ABA) levels. To further elucidate the regulatory mechanism, eight genes for GA biosynthesis and signaling, including Ps CPS, Ps KS, Ps GA_3 ox, Ps GA2 ox, Ps GID1 b, Ps GID1 c, Ps DELLA, and Ps GID2 were cloned for the first time, and sequence analysis was also performed. The results suggested that all the cloned genes have conserved structure as each homologous gene reported in the other species. Phylogenetic trees constructed by the each cloned gene showed that the phylogenetic evolutionary relationship of P. suffruticosa was closely related to Vitis vinifera. The expression patterns of the above genes, and genes for ABA and IAA biosynthetic and signaling, and the flowering time were also investigated. Most of the above genes showed higher expression in the control buds than those in the GA_3 treated buds at six developmental stages, whereas the expression levels of PsSOC1 and PsSPL9 were up-regulated by GA_3 treatment. The results also showed that the GA-biosynthetic and signaling pathways are conserved in tree peony, and the PsCPS, PsGA_3 ox, PsGA2 ox, PsGID1, PsDELLA, and PsGID2 genes are necessary for feedback regulation of GAs. Furthermore, hormone changes promoted PsSOC1 and PsSPL9 expression, and repressed PsSVP expression, which contributed to the improvement flowering quality in tree peony of forcing culture.     

不同施氮量与种植密度对水稻穗上不同部位结实特性的影响

以优质食味水稻品种南粳9108为材料,研究不同施氮量与种植密度下水稻穗上不同部位结实特性的差异。结果表明,施氮量与种植密度对水稻的结实特性有一定的互作效应,低种植密度下,适当增加氮肥可降低水稻籽粒的空粒率和秕粒率。从水稻穗位上看,稻穗一次枝梗的结实特性优于二次枝梗,稻穗不同部位和一、二次枝梗不同部位的结实特性均表现为上部>中部>下部。在相同的一次或二次枝梗上不同粒位的空秕粒率一般以第2粒最高,二次枝梗不同粒位的空秕粒率以第1粒为最低,第2,3粒最高。     

Molecular identification and enzymatic properties of laccase2 from the diamondback moth Plutella xylostella (Lepidoptera: Plutellidae)

Laccase(EC 1.10.3.2)is known to oxidize various aromatic and nonaromatic compounds via a radical-catalyzed reaction,which generally includes two types of laccase,Lac1 and Lac2.Lac1 oxidizes toxic compounds in the diet,and Lac2 is known to play an important role in melanizing the insect exoskeleton.In this study,we cloned and sequenced the cDNA of the diamondback moth,Plutella xylostella Lac2(PxLac2),from the third instar larvae using polymerase chain reaction(PCR)and rapid amplification of cDNA ends techniques.The results showed that the full-length PxLac2 cDNA was 1 944 bp long and had an open reading frame of 1 794 bp.PxLac2 encoded a protein with 597 amino acids and had a molecular weight of 66.09 kDa.Moreover,we determined the expression levels of PxLac2 in different stages by quantitative PCR(qPCR).The results indicated that PxLac2 was expressed differently in different stages.We observed the highest expression level in pupae and the lowest expression level in fourth instar larvae.We also investigated the enzymatic properties of laccase,which had optimal activity at pH 3.0 and at 35°C.Under these optimal conditions,laccase had a Michaelis constant(K_m)of 0.97 mmol L~(-1),maximal reaction speed(V_m)of 56.82 U mL~(-1),and activation energy(E_a)of 17.36 kJ mol~(-1) to oxidize2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid ammonium salt).Type II copper enhanced laccase activity below 0.8mmol L~(-1) and reduced enzyme activity above 0.8 mmol L~(-1) with an IC_(50) concentration of 1.26 mmol L~(-1).This study provides insights into the biological function of laccase.     

氮素水平对水稻产量与品质的影响

在大田试验条件下,研究不同施氮量对中籼稻汕优63和中粳稻武育粳3号稻米品质及产量的影响。结果表明:施氮量对稻米的出糙率、精米率、粒长宽比、透明度和直链淀粉含量无显著影响。稻米蛋白质含量随施氮量的增加而增加。汕优63、武育粳3号施氮(纯氮)量分别为0~16和0~24 g.m-2,穗数、每穗粒数、产量、整精米率、最高黏度和崩解值随施氮量的增加而增加,稻米的垩白粒率、垩白度和消减值则随施氮量的增加而降低。汕优63、武育粳3号施氮(纯氮)量分别超过16、24 g.m-2时,结实率、千粒重、产量、整精米率、最高黏度和崩解值随施氮量的增加而降低,稻米的垩白粒率、垩白度和消减值则随施氮量的增加而增加。在该试验条件下,可将施氮(纯氮)量160和240 kg.hm-2分别作为籼稻汕优63和粳稻武育粳3号高产优质的氮肥施用量指标。     

Evaluating the efficacy of an attenuated Streptococcus equi ssp. zooepidemicus vaccine produced by multi-gene deletion in pathogenicity island SeseCisland_4

Streptococcus equi ssp. zooepidemicus(SEZ) is a pathogen associated with a wild range of animal species. Frequent outbreaks have occurred in recent years in pigs, horses, goats and dogs which is liable to infect humans. There is a lack of efficient vaccines against this disease and the occurrence of antibiotic resistance may render drug therapies ineffective. In this study, gene deletion mutant(ΔSEZ) in pathogenicity islands SeseCisland_4 was constructed. The mutant ΔSEZ had a 52-fold decrease in 50% lethal dose(LD_(50)) and had less capacity to adhere epithelial cells. Importantly, immunization of mice with attenuated vaccine ΔSEZ at the dose of 10~2 colony-forming units(CFU) mL~(–1) elicited a significant humoral antibody response, with an antibody titer of 1:12 800. Therefore, 10~2 CFU mL~(–1) might be used as the appropriate immune dose for the attenuated vaccine ΔSEZ, which provided mice with efficient protection against virulent SEZ. In addition, the hyperimmune sera against 10~2 CFU m L–1 attenuated vaccine ΔSEZ could confer significant protection against virulent SEZ infection in the passive immunization experiment and exhibited efficient bactericidal activity in the whole blood assay. Meanwhile, no viable bacteria was detected in blood when mice were immunized with ΔSEZ at the dose of 10~2 CFU mL~(–1) via hypodermic injection. Thereafter, the mutant ΔSEZ at the dose of 10~2 CFU mL~(–1) could confer significant protection in mice and had less negative effects on host, which could be an effective attenuated vaccine candidate for the prevention of SEZ.