西北地区苹果褐斑病数学预测模型建立与分析

依据2001～2010年我国西北苹果主产区的气象数据与苹果褐斑病(Marssonina coronaria)病害发生流行数据,构建了平均气温(T)、相对湿度(Hm)、风速(Ws)及地表温度(St)影响下苹果褐斑病的流行趋势预测模型,并利用多元回归分析方法建立了苹果褐斑病预测模型.结果表明环境因素严重影响苹果褐斑病的发生和流行,苹果褐斑病预测模型为:f(T,Hm,Ws,St)=1.317T+0.002Hm+0.047Ws+0.001St-11.885[f(T,Hm,Ws,St)为病情指数].苹果褐斑病发生的T为14℃～15℃,Hm为45%,Ws为13m/s,St为10℃.大发生条件为T为20℃～23℃,Hm在70%～90%之间,Ws为0m/s～2m/s及St为22°.     

低氧应激下甘肃鼢鼠与SD 大鼠心脏抗氧化酶和ATP 酶活性的比较

为探讨低氧应激下甘肃鼢鼠心脏对抗氧化损伤和电生理紊乱的可能机制,对甘肃鼢鼠和SD 大鼠在4. 5%氧浓度下分别进行2 h、4 h、6 h、8 h、10 h、16 h 低氧应激,比较常氧和各时程低氧下二者心脏超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽还原酶(GR)和Ca^{2 +} - ATP 酶、Ca^{2 +} - Mg^{2 +} - ATP 酶、Na ⁺ - K ⁺ - ATP酶活性,以丙二醛(MDA)含量作为机体氧化损伤指标。结果显示,常氧组甘肃鼢鼠GR 活性比SD 大鼠高,SOD、CAT、Ca^{2 +}-ATP 酶、Ca^{2 +} - Mg^{2 +} - ATP 酶和Na ⁺ - K ⁺ - ATP 酶活性及MDA 含量与SD 大鼠相比均无显著性差
异;低氧组甘肃鼢鼠SOD、CAT、GR、Ca^{2 +} - ATP 酶、Ca^{2 +} - Mg^{2 +} - ATP 酶和Na ⁺ - K ⁺ - ATP 酶活性迅速升高,显著高于SD 大鼠,MDA 含量则显著低于SD 大鼠。说明甘肃鼢鼠心脏通过提高抗氧化酶活性清除低氧诱导产生的多余自由基,并通过提高ATP 酶活性保证心电活动正常、心率稳定,应对低氧应激。     

红水河暗色唇鲮的年龄与生长研究

为了解广西壮族自治区红水河暗色唇鲮Semilabeo obscurus的年龄结构及生长特性,于2022年11月在红水河采集71尾样本。结果显示:样本群体由1～6龄组成,优势年龄组为3龄,占总数的39.44%;平均体长16.51 cm,平均体重122.35 g,体重(W)与体长(L)呈幂函数相关:♀:W=0.039L^2.827;♂:W=0.025L^2.963;群体:W=0.026L^2.964,属匀速生长型;生长特性符合von Betalanffy生长方程:♀:L_t=24.74[1-e⁽-0.4195(t+0.2438))],W_t=340.65[1-e⁽-0.4195(t+0.2438))]^2.827;♂:L_t=26.70 [1-e⁽-0.3103(t+695 1))],W_t=427.98[1-e⁽-0.3103(t+0.695 ...     

基于控制试验的植株数量及空气温湿度与负离子的关系

空气负离子(NAI)作为评价大气环境质量的重要指标,以往研究侧重于野外观测试验,关于控制条件下的NAI研究甚少.本研究于2018年9-10月在浙江钱江源森林生态站杭州虎山试验基地人工气候室内控制相同气候条件下,对枫香和红豆杉不同植株数量的NAI浓度进行连续定位监测;在其他条件一致的情况下,分别设置空气温度和相对湿度的不同梯度,监测NAI浓度变化,旨在为探究森林植被和气象因子对NAI作用机理提供理论依据.结果表明: 两种植物显著提高NAI浓度.植株数量与NAI浓度呈极显著正相关,二者在0~50株范围内最符合二次函数关系,枫香、红豆杉的NAI浓度与株数的拟合方程分别为y=-0.0484x²+4.7005x+345.7(R²=0.62)、y=-0.0207x²+1.9189x+365.91(R²=0.34).NAI浓度与空气温度在5~30 ℃范围内呈极显著正相关,拟合方程为y=0.4139x²-9.2229x+89.919(R²=0.92).NAI浓度与空气相对湿度在56％~87％范围内呈极显著正相关,拟合方程为y=3.6508e^{0.0526x(R2=0.94).}     

奶牛乳铁蛋白基因5非翻译区PCR­SSCP多态性分析

采用PCR-SSCP技术,对奶牛乳铁蛋白基因5^'非翻译区1122bp序列进行多态性分析。在该区所划分的5个亚片段上,发现 Blf 5^'-1(227bp),Blf 5^'-3(175bp)和Blf 5^'-5（293bp）3个DNA片段存在多态。进一步对这3个片段进行测序分析,在Blf 5^'-1序列中,发现位于转录起始位点上游-926和-915位分别有G→A及T→G点突变;在Blf 5^'-3片段中,-478位存在G的插入;Blf 5^'-5位点上,发生-28位的C颠换为A和+33位的G颠换为C两处突变。利用TFSEARCH （ver.1.3）软件对乳铁蛋白基因5^'非翻译区潜在调控元件及蛋白质结合位点进行了预测,结果显示5^'非翻译区的突变引起了蛋白质结合因子的变化。     

闽江口海蜇渔业生态学研究

根据1993年5～9月的调查材料,研究了福建闽江口海域海蜇的渔业生态学。其密度和生物量高峰分别出现在6月20日和7月10日;群体伞径范围为18～546mm,平均328.8mm.体重范围为0.5～9540g,平均2877.4g;伞径为345～485mm的雌性有性繁殖力在1120.6×10⁴～3754.8×10⁴粒,平均2444.7×10⁴粒,有性生殖期在8月初至11月;采用高次方程和指数高次方程分别模拟伞径和体重生长,生长方程为:Φ_t=12.1337+17.7048t+3.1385t²-0.2049t³+0.00302t⁴,log_eW_t=-0.5749+1.4818t-0.0771t²+0.00129t³;以生物经济学原理确定7月20日为合理开捕期,开捕伞径为465.8mm.讨论分析了执行合理开捕期和开捕规格,对保护和合理利用海蜇资源,提高经济效益的重要意义.     

硒的价态与浓度对香橙幼苗生长和AsA-GSH循环的影响

以盆栽香橙为试材,分析不同施用浓度Se⁶⁺和Se⁴⁺对植株生长和抗坏血酸(AsA) -谷胱甘肽(GSH)循环的影响.结果表明: 两种价态硒均可促进香橙生长,主要表现在增加了叶面积、株高、鲜质量和干质量.施用Se⁶⁺显著提高了香橙硒含量,且硒主要分配在叶片;而施用Se⁴⁺虽能提高硒含量,但其含量远低于Se⁶⁺处理,且主要分配在根系.施硒提高了叶片叶绿素和过氧化氢(H₂O₂)含量,且Se⁶⁺处理高于Se⁴⁺处理.Se⁶⁺浓度≤2.0 mg·L^-1处理能提高谷胱甘肽还原酶(GR)与谷胱甘肽过氧化物酶(GPX)活性、GSH与氧化型谷胱甘肽(GSSG)量,Se⁶⁺浓度≥4.0 mg·L^-1处理降低GSH循环的物质和酶活性;而Se⁴⁺浓度≤ 2.0 mg·L^-1处理能提高脱氢抗坏血酸还原酶(DHAR)与抗坏血酸过氧化物酶(APX)活性,且具有较高的AsA/[AsA+脱氢抗坏血酸(DHA)]比值,Se⁴⁺浓度≥4.0 mg·L^-1 处理的GSH循环的物质和酶活性升高.综上,硒的不同价态和施用浓度对AsA-GSH循环相关物质含量和酶活性的影响不同.结合香橙生长指标和抗氧化水平,Se⁶⁺和Se⁴⁺的适宜浓度分别为2.0和4.0 mg·L^-1.     

广州市酸雨成分及其相关分析

分析了2003年11月-2004年10月广州市龙洞79次降雨的化学组分及其各组分之间的相关关系。得出:(1)观测期间总降雨量1359.8,单次降雨的pH值在3.22-7.29之间,平均4.83;以pH<5.6为酸雨的临界值,则酸雨频率占降雨次数的77.2%或占降雨量的81.5%;(2)广州降水的化学组成以SO₄^2-、Cl^-和NO₃^-为主要阴离子,阳离子中以Ca²⁺、NH₄⁺、Na⁺所占比例较大;(3)广州地区酸雨属硫酸型,降雨中的SO₄^2-、NO₃^-、PO₄^3-、Cl^-、NH₄⁺、K⁺、Na⁺、Ca²⁺、Mg²⁺离子浓度分别为0.150,0.045,0.002,0.163,0.058,0.006,0.013,0.083和0.007mol·L^-1,单次降雨的SO₄^2-/NO₃^-比值在0.42-17.6之间,年平均为3.33,该值比西南地区相应值低许多,而且与同一地点5年前测定的结果相比较,SO₄^2-/NO₃^-比值有变小的趋势,说明广州地区NO_x排放量高逐渐增加;(4)广州地区酸雨有着明显的季节变化规律,春、夏、秋季酸雨比较严重,冬季酸雨较少出现,这与西南地区的重庆(冬季酸雨较春季严重)酸雨季节规律相反;(5)通过SPSS统计分析得出,雨量与各化学组分含量成负相关,说明雨量对大气污染物质的稀释作用;酸雨各化学成分之间存在着一定的相关性,如SO₄^2-与Ca²⁺,NO₃^-与NH₄⁺,K⁺与Na⁺、Mg²⁺离子,Na⁺与Mg²⁺等都存在显著的正相关,利用这种相关可以建立某些酸雨成分的预测模型。     

遗传学实验

大肠杆菌的杂交与致育因子F有关。有F因子的 细菌称为F⁺,没有F因子的细菌称为F^-。一般F^-菌 株之间不能进行杂交。F因子有两种状态— t游离状 态和整合状态（F因子插人到染色体的一定位置上,所 以F因子是附加体）,前者称F⁺菌株,后者称为高频重 组或Hfr.F⁺和Hfr都能与F-菌株进行杂交,但重组矫 率不同,Hfr比F⁺高1,000倍,一般为10-^-3-10^-4.     

α1-肾上腺素受体对胶原诱导性关节炎小鼠Treg细胞功能的影响*

目的: 利用胶原诱导性关节炎 (CIA) 模型小鼠,探讨去甲肾上腺素 (NE) 及其α1-肾上腺素受体 (AR ) 对CIA小鼠Treg细胞的作用。方法: 雄性DBA/1小鼠 32 只,随机分为对照组 (n=8) 和CIA模型组 (n=24)。II型胶原 (CII) 乳剂100 μl 尾根部注射DBA/1小鼠制备CIA小鼠模型,在初次免疫后第 41 日,用免疫荧光法检测小鼠脾脏中CD4⁺T与α1-AR的共定位情况;用Western blot法检测小鼠踝关节和脾脏中α1-AR的蛋白表达。分离纯化CIA小鼠脾脏中CD4⁺ T细胞,用抗CD3和抗CD28的单克隆抗体刺激CD4⁺T细胞,进行细胞培养,分为未加药组和加药组,加药组用NE或α1-AR激动剂苯肾上腺素 (phenylephrine) 处理细胞,用流式细胞术检测CIA小鼠CD4⁺T细胞中Treg的细胞数;用Western blot法检测CIA小鼠CD4⁺T中转化生长因子-β (TGF-β) 和IL-10的蛋白表达。结果: CD4⁺ T细胞能够表达α1-AR;与对照组相比,CIA小鼠踝关节和脾脏中α1-AR的蛋白表达显著降低(P＜0.01);与未加药的CIA小鼠的CD4⁺ T细胞相比,NE加入后的CIA小鼠CD4⁺ T细胞中Treg细胞的功能显著增强(P＜0.01);α1-AR激动剂phenylephrine加入后的CIA小鼠CD4⁺ T细胞中Treg细胞的功能显著增强(P＜ 0.01)。结论: 激活CIA小鼠CD4⁺ T细胞上的α1-AR可增强Treg细胞的功能,促进CD4⁺ T细胞向Treg细胞方向分化,发挥抗炎作用。     

Ectopic dermal ridge configurations on the interdigital webbings and postaxial marginal portion of the hindlimb in Hammertoe mutant mice (Hm)

The effects of the hereditary malformation of Hammertoe mutant mice (gene symbol Hm) on the digital pads and dermal ridge configurations on their hindlimbs were examined. In the wild‐type (+/+) mice with normally separated digits, dermal ridges developed only on the pads. Heterozygous (Hm/+) and homozygous (Hm/Hm) mutant mice, however, had a broad big toe, fused interdigital soft tissues, reduced claws, an extra rudimentary postaxial digit and camptodactyly. The dermal ridges appeared not only on the pads, affected in their number and configurations, but also on the ventral surface of the interdigital webbings and postaxial marginal area exhibiting an extra rudimentary digit and webbing. These aberrant configurations may be related to the abnormal occurrence of programmed cell death (PCD) in the interdigital zones and the postaxial marginal portion in Hm/+ and Hm/Hm mice. That is, the diminished cell death may fail to decrease the cell density in the interdigital zones and postaxial marginal portion and result in the webbing and an extra rudimentary digit and webbing, respectively. Simultaneously, it could also interrupt the migration of surviving cells of these areas toward the neighboring digits and the distal area of the sole and produce the ectopic dermal ridges on the way to the as yet unformed (presumptive) digital and plantar volar pads. The present findings suggest that normal interdigital and pre/postaxial PCD contributes not only to the separation of digits, the initial formation of individual digits of different sizes, and the inhibition of the extra digit but also to the development of the presumptive digital and plantar pads, including dermal ridges. J. Morphol., 2008. © 2008 Wiley‐Liss, Inc.     

Assessment of real-time PCR for quantification of Legionella spp. in spa water

Aim: To identify media and environmental conditions suitable for rapid mycelial growth and sporulation of Diplocarpon mali. Methods and Results: Liquid shake cultures were used to evaluate effects of media and environmental conditions on mycelial growth and conidial production of D. mali. Carrot sucrose broth (CSB), potato and carrot dextrose broth (PCDB) and potato and carrot sucrose broth (PCSB) were most favourable for rapid mycelial growth. PCDB, PCSB, PCB (potato and carrot broth) and carrot dextrose broth (CDB) were favourable for conidial production. All carbon sources tested and peptone favoured for mycelial growth. Carbon and nitrogen sources tested did not significantly stimulate conidial production. The optimum temperature for mycelial growth and conidial production was 25°C. No mycelial growth occurred at 5 or 30°C, but D. mali survived at these temperatures. Active mycelial growth occurred at pH 5–7, and pH 5–8 was favourable for sporulation. Conclusions: PCDB and PCSB incubated at 25°C for 14 day are recommended for mycelial growth and conidial production of D. mali. Significance and Impact of the Study: The information generated in this study will facilitate mycological and pathological research on D. mali and Marssonina leaf blotch of apple caused by D. mali.     

盘二孢属一新种——花椒盘二孢

花椒盘二孢Marssonina zanthoxyla Y.J.Lu et G.L.Li sp.nov.是寄生在芸香科花椒属植物上的一种寄生菌,本文有汉文和拉丁文的形态描述和新种的讨论。     

Ecological optima and tolerances of Thelypteris limbosperma,Athyrium distentifolium,and Matteuccia struthiopteris along environmental gradients in Western Norway

The distribution and abundance of Thelypteris limbosperma, Athyrium distentifolium, and Matteuccia struthiopteris are modelled statistically in relation to 14 environmental variables along the major climatic, topographic, and edaphic gradients in western Norway. The data are from 624 stands from which measurements or estimates of mean January and mean July temperatures, humidity, altitude, aspect, and slope are available. From 182 of these stands eight soil variables have also been measured. The species responses are quantified by two numerical methods: Gaussian logit regression and weighted averaging (WA) regression. The estimated WA optima suggest that A. distentifolium has an ecological preference for low July and January temperatures, high altitudes, and soils of low-medium pH and base content. The species shows statistically significant Gaussian responses with summer temperature, humidity (= Martonnes humidity index), altitude, slope, aspect, pH, cation exchange capacity, and base saturation with optima of 8.7 °C, 188.9, 1220 m, 28°, 29°, 4.8, 13.77 mEq 100 g dry soil^-1, and 13.4%, respectively. These suggest that the occurrence and relative abundance of A. distentifolium are well predicted by summer temperature, topography, and soil pH and base status. T. limbosperma has WA optima that suggest that it favours moderately high winter and summer temperatures, high humidity, medium altitude, and soils of low pH and base content. It has significant Gaussian responses to summer temperature (optimum =12.6 °C), winter temperature (-1.8 °C), humidity (179.2), altitude (459.5 m), slope (22.5°), and Na (0.7 mg 100 g dry soil^-1). These suggest that climatic factors, altitude, and slope are significant predictors for its occurrence and abundance. M. struthiopteris has high WA optima for summer temperature, pH, Ca, Mg, K, Na, cation exchange capacity (CEC), and base saturation, and a low optima for humidity and winter temperature. Of these, summer temperature (16.0 °C), Ca (63.1 mg 100 g dry soil^-1), Mg (41.0 mg 100 g dry soil^-1), K (23.6 mg 100 g dry soil^-1), Na (5.0 mg 100 g dry soil^-1), CEC (60.7 mEq 100 g dry soil^-1), and base saturation (56.3%) have significant Gaussian logit responses, as do aspect (150.2°) and loss-on-ignition (9.4%). These results suggest that the occurrence and relative abundance of M. struthiopteris are well predicted by high soil base cations, a generally southern aspect, low organic content in the soil, and high July temperatures.     

<Emphasis Type="Italic">Agrobacterium</Emphasis>-mediated transformation of Perilla (<Emphasis Type="Italic">Perilla frutescens</Emphasis>)

Agrobacterium tumefaciens-mediated transformation system for perilla (Perilla frutescens Britt) was developed. Agrobacterium strain EHA105 harboring binary vector pBK I containing bar and γ-tmt cassettes or pIG121Hm containing nptII, hpt, and gusA cassettes were used for transformation. Three different types of explant, hypocotyl, cotyledon and leaf, were evaluated for transformation and hypocotyl explants resulted in the highest transformation efficiency with an average of 3.1 and 2.2%, with pBK I and pIG121Hm, respectively. The Perilla spp. displayed genotype-response for transformation. The effective concentrations of selective agents were 2 mg l⁻¹ phosphinothricin (PPT) and 150 mg l⁻¹ kanamycin, respectively, for shoot induction and 1 mg l⁻¹ PPT and 125 mg l⁻¹ kanamycin, respectively, for shoot elongation. The transformation events were confirmed by herbicide Basta spray or histochemical GUS staining of T₀ and T₁ plants. The T-DNA integration and transgene inheritance were confirmed by PCR and Southern blot analysis of random samples of T₀ and T₁ transgenic plants.     

Detailed description of developmental growth stages of Diplocarpon rosae in Rosa: a core building block for efficient disease management

Black spot disease of roses caused by the ascomycetous fungus Diplocarpon rosae teleomorph (Marssonina rosae anamorph) is a widespread and devastating disease. Despite considerable progress in the management of black spot disease in the recent years, it is still unclear by which mechanisms this fungus colonises and invades the host system, and without a good knowledge of such infection machineries, it not possible to fully overcome the challenges of D. rosae infection. By exploring research contributions up to date, we highlight in this review the ultrastructure of D. rosae infection cycle in the host cell by emphasising on several aspects related to its in vitro and in vivo germinations, the infection mechanisms of the fungus, the different fungal structures formed in the host cells and the optimum storage conditions for D. rosae to retain its viability and pathogenicity over time. Here, attention is particularly focused on the asexual life cycle of D. rosae, with the sexual cycle being briefly discussed. In addition, a new dimension of research approaches to effectively control black spot disease of roses, that is, how to accurately use the advanced biotechnology tools to speed up the current state of the disease management, is proposed here.     

Isolation of Trichoderma and Evaluation of their Antagonistic Potential against Alternaria porri

Nine isolates of Trichoderma were collected from Assiut Governorate, Egypt, as leaf surface and endophytic fungi associated with onion flora stalks. Four isolates were identified as Trichoderma harzianum, while five isolates were belonging to Trichoderma longibrachiatum. The antagonistic activity of these isolates against onion purple blotch pathogen Alternaria porri was studied in vitro using dual culture assay. All tested Trichoderma isolates showed mycoparasitic activity and competitive capability against the mycelial growth of A. porri. Mycoparastic activity of Trichoderma was manifested morphologically by the overgrowth upon the mycelial growth of the pathogen and microscopically by production of coiling hyphae around pathogen hyphae. Isolates of T. harzianum exhibited high ability to compete on potato dextrose agar (PDA) medium causing the maximum rate of pathogen inhibition (73.12%), while isolates of T. longibrachiatum showed inhibition rate equalling 70.3%. Chitinase activity of Trichoderma was assayed, and T. harzianum Th‐3013 showed the maximum value contributing 2.69 U/min. Application of T. harzianum Th‐3013 to control purple blotch disease in vivo under greenhouse conditions caused disease reduction up to 52.3 and 79.9% before and after 48 h of pathogen inoculation, respectively, while the fungicide Ridomil Gold Plus caused disease reduction comprising 56.5 and 71.7%, respectively. This study proved that T. harzianum Th‐3013 as a biocontrol agent showed significant reduction in onion purple blotch disease compared with the tested fungicide.     

Apoptosis of CD4+CD25high T Cells in Type 1 Diabetes May Be Partially Mediated by IL-2 Deprivation

Background

Type 1 diabetes (T1D) is a T-cell mediated autoimmune disease targeting the insulin-producing pancreatic β cells. Naturally occurring FOXP3⁺CD4⁺CD25^high regulatory T cells (T_regs) play an important role in dominant tolerance, suppressing autoreactive CD4⁺ effector T cell activity. Previously, in both recent-onset T1D patients and β cell antibody-positive at-risk individuals, we observed increased apoptosis and decreased function of polyclonal T_regs in the periphery. Our objective here was to elucidate the genes and signaling pathways triggering apoptosis in T_regs from T1D subjects.

Principal Findings

Gene expression profiles of unstimulated T_regs from recent-onset T1D (n = 12) and healthy control subjects (n = 15) were generated. Statistical analysis was performed using a Bayesian approach that is highly efficient in determining differentially expressed genes with low number of replicate samples in each of the two phenotypic groups. Microarray analysis showed that several cytokine/chemokine receptor genes, HLA genes, GIMAP family genes and cell adhesion genes were downregulated in T_regs from T1D subjects, relative to control subjects. Several downstream target genes of the AKT and p53 pathways were also upregulated in T1D subjects, relative to controls. Further, expression signatures and increased apoptosis in T_regs from T1D subjects partially mirrored the response of healthy T_regs under conditions of IL-2 deprivation. CD4⁺ effector T-cells from T1D subjects showed a marked reduction in IL-2 secretion. This could indicate that prior to and during the onset of disease, T_regs in T1D may be caught up in a relatively deficient cytokine milieu.

Conclusions

In summary, expression signatures in T_regs from T1D subjects reflect a cellular response that leads to increased sensitivity to apoptosis, partially due to cytokine deprivation. Further characterization of these signaling cascades should enable the detection of genes that can be targeted for restoring T_reg function in subjects predisposed to T1D.     

A Two-Component DNA-Prime/Protein-Boost Vaccination Strategy for Eliciting Long-Term,Protective T Cell Immunity against Trypanosoma cruzi

In this study, we evaluated the long-term efficacy of a two-component subunit vaccine against Trypanosoma cruzi infection. C57BL/6 mice were immunized with TcG2/TcG4 vaccine delivered by a DNA-prime/Protein-boost (D/P) approach and challenged with T. cruzi at 120 or 180 days post-vaccination (dpv). We examined whether vaccine-primed T cell immunity was capable of rapid expansion and intercepting the infecting T. cruzi. Our data showed that D/P vaccine elicited CD4⁺ (30-38%) and CD8⁺ (22-42%) T cells maintained an effector phenotype up to 180 dpv, and were capable of responding to antigenic stimulus or challenge infection by a rapid expansion (CD8>CD4) with type 1 cytokine (IFNγ⁺ and TFNα⁺) production and cytolytic T lymphocyte (CTL) activity. Subsequently, challenge infection at 120 or 180 dpv, resulted in 2-3-fold lower parasite burden in vaccinated mice than was noted in unvaccinated/infected mice. Co-delivery of IL-12- and GMCSF-encoding expression plasmids provided no significant benefits in enhancing the anti-parasite efficacy of the vaccine-induced T cell immunity. Booster immunization (bi) with recombinant TcG2/TcG4 proteins 3-months after primary vaccine enhanced the protective efficacy, evidenced by an enhanced expansion (1.2-2.8-fold increase) of parasite-specific, type 1 CD4⁺ and CD8⁺ T cells and a potent CTL response capable of providing significantly improved (3-4.5-fold) control of infecting T. cruzi. Further, CD8⁺T cells in vaccinated/bi mice were predominantly of central memory phenotype, and capable of responding to challenge infection 4-6-months post bi by a rapid expansion to a poly-functional effector phenotype, and providing a 1.5-2.3-fold reduction in tissue parasite replication. We conclude that the TcG2/TcG4 D/P vaccine provided long-term anti-T. cruzi T cell immunity, and bi would be an effective strategy to maintain or enhance the vaccine-induced protective immunity against T. cruzi infection and Chagas disease.