Effects of the relationship between area under disease progress curve of angular leaf spot and yield of seed cotton in northeastern Nigeria

Bacterial blight induced by Xanthomonas campestris pv. malvacearum causes substantial yield losses of cotton in Nigeria annually. A field study was carried out for two seasons to investigate the reaction of a selection of 10 different cotton genotypes to bacterial blight and to examine the relationship between the area under disease progress curve (AUDPC) and the yield of seed cotton in the northeastern cotton region of Nigeria. All 10 genotypes reacted positively to the disease with SAMCOT-13 having the highest incidence particularly at Dowaya and Kem locations during the two seasons. This investigation also revealed that SAMCOT-13 and Ex-Benin had the highest AUDPC while TX-CDP37HH-1-83, TAMCOT CAMD-E and TAMCOT SP-21S had the lowest AUDPC indicating relative susceptibility and resistance, respectively. However, it was observed that TX-CDP37HH-1-83, SAMCOT-11 and Ex-Benin had the highest mean yields of 1441.5 kg/ha and 1494.7 kg/ha at Dowaya, Kem and Ngurore, respectively. The results also show a highly significant (P = 0.01) negative correlation between AUDPC and yield in the three locations. These findings suggest that TX-CDP37HH-1-83, TAMCOT CAMD-E and TAMCOT SP-21S are moderately resistant to the disease in view of their lower AUDPC and are considered promising.     

Distribution and Races of Xanthomonas axonopodis pv. malvacearum on Cotton (Gossypium hirsutum) in Uganda

Surveys in 1995 and 1996 showed that bacterial blight caused by Xanthomonas axonopodis pv. malvacearum occurs throughout the main cotton growing areas of Uganda, causing seedling blight, angular leaf spot and bacterial boll rot. During the vegetative and early fruiting stages of crop growth, severe symptoms of `blackarm' spread from leaves to the stem, causing loss of fruiting branches. A set of Upland cotton cultivars ( Gossypium hirsutum ) were then used to determine the races of the blight bacterium present in Uganda. Many of the isolates induced moderate to severe symptoms on all the test hosts except 101–102B, indicating infection with race 10 or 18. The next most common isolate was race 7. Races 16 and 6 were also identified and 23% of isolates caused symptoms on all the differential cultivars including 101–102B, results indicating the presence of a race of the pathogen which may be the same as that identified in countries neighbouring Uganda and designated as race 20.     

棉铃虫的取食营养特点与棉花抗虫素分布的关系

通过研究棉花8个部位对棉铃虫Helicoverpa armzgera的营养效果和其所含次生物质类萜烯和单宁的浓度及在组织中的分布,揭示了棉铃虫取食营养特点与棉花次生化学的关系。棉铃虫取食顶尖、转移蛀食蕾铃的习性,与有关器官或组织对幼虫的营养效果密切相关,而营养效果主要取决于类萜烯和单宁的含量。棉花顶尖嫩叶中单宁浓度随着棉花的生长发育呈升高的趋势;类萜烯浓度在第四真叶期、第六真叶期和现蕾初期之间出现一个明显的底谷,而此期幼虫主要为害顶尖。蕾铃外层的苞叶、花萼、花瓣和铃皮,因次生物质含量高,不利于幼虫生长,相反内部的花粉、子房和铃心,次生物质含量低,营养效果好,顶尖嫩叶则介于其间。类萜烯存在于组织的色素腺内,分布集中;单宁则散布于组织中,偏多分布于组织外层。结果证实,昆虫在寄主植物上的取食方式是昆虫对寄主体内变动的次生化学的一种适应,它使昆虫付出尽少代价获得最适营养效果。     

转ACO2基因优质棉生长特性及其对田间昆虫群落的影响

新型转基因棉花的生长特性及其对棉田昆虫群落的影响是转基因棉花环境安全评价的重点内容,也是转基因棉花环境安全评价过程必须评价的内容。选用新型转ACO2基因棉花及其亲本中棉所24为试验材料,于2013年和2014年在河南安阳进行了田间棉花生长势及其对昆虫群落影响的试验。在棉花苗期、蕾期、花铃期和铃期,转基因棉花叶片干重与其亲本均无显著差异,苗期、蕾期叶面积和比叶面积与亲本无显著差异,花铃期、铃期叶面积与比叶面积均显著高于亲本材料,其中2013年分别比对照高27.1%和26.2%,2014年分别高16.0%和19.9%,差异均达显著水平;株高、主茎叶片数、大铃数、小铃数和蕾铃脱落数均与亲本材料相当,但铃期果枝数显著高于亲本材料;3个棉铃虫(Helicoverpa armigera Hubner)发生高峰期,转优质基因棉花对棉铃虫和甜菜夜蛾幼虫校正死亡率与亲本对照材料无显著差异;昆虫群落、害虫亚群落和天敌亚群落昆虫个体总数均显著高于亲本棉田,两年分别比对照高11.5%、10.5%、33.4%和12.0%、8.1%、63.1%,其中2013年差异达显著水平,2014年由于昆虫发生程度总体较轻,差异不显著,但所属"目"、"科"、"种’、生物多样性指数、优势种昆虫及其优势度与其亲本棉田无显著差异,表明外源基因ACO2导入后,对棉田昆虫发生及其群落结构没有影响,棉花生长势无明显优势,但产量构成性状在部分时期增强。上述研究初步明确了新型转基因棉花生存竞争势态和棉田昆虫群落发生规律与动态,可为新型转基因棉花环境安全评价提供理论依据,同时为转基因棉花环境安全评价积累科学数据。     

Spectrum of biocontrol bacteria to control leaf,root and vascular diseases of dry bean

Bacterial biocontrol agents, previously selected to control bacterial blight and anthracnose, were evaluated for the control of bacterial wilt, fusarium wilt, charcoal rot and angular leaf spot of dry beans. The seeds were microbiolized with these bacteria singly or in combinations. The microbiolization resulted in reduction of severity of all four diseases, showing wide spectrum of diseases control by these bacteria. However, the severity reduction of all four diseases by combination C01 composed of isolates DFs093 and DFs769 of Bacillus cereus and DFs831 of Pseudomonas fluorescens was significantly higher than if the seeds were microbiolized by a single isolate of a biocontrol agent.     

Fungi associated with boll and lint rot of cotton in Southern Khorasan province of Iran

During surveys of cotton fields since 2009 in the Southern Khorasan province of Iran, symptomatic samples with lint and boll rot disease signs were collected and cultured on potato dextrose, malt extract agar and PARPH media. Several fungal isolates were obtained and identified. Among them, we found Penicillium expansum which to our knowledge is reported as a new pathogen of cotton worldwide; Fusarium semitectum and Nigrospora oryzae (associated to mite Siteroptes sp.) were also recorded as lint or boll rot pathogens for the first time from Iran. Disease symptoms and fungal characteristics of Exserohilum rostratum, which has been reported briefly as a new causal agent of boll and lint rot disease, are illustrated and presented. In addition, Aspergillus niger and Rhizopus spp. were also isolated frequently from lint parts of cotton prior to harvesting. There was a positive correlation between disease occurrence and spiny cotton bollworm damage. The pathogenicity test was characterised by inoculating the attached and detached bolls of cotton plants.     

Pathogenic aspects of Pantoea agglomerans in relation to cotton boll age and Dysdercus cingulatus (Fabricius) transmitting seed and boll rot in cotton germplasm

Bacterial seed and boll rot is a newly emerging cotton disease in Pakistan. Twenty-one cotton varieties were screened to find resistance source against the disease. None of these was found to be resistant. Five cotton varieties (CIM-595, MK2, BT-986, BT-986 & SG-1) having 700–1400 Area under disease progress curve (AUDPC) units were found to be moderately resistant to the disease. SLH-317, FH-942, BT-222, BT-666, MNH-457 ranging from 1401–1700 AUDPC units were moderately susceptible while MNH-456, SLH-336, 9811, FH-942, MNH-886 susceptible to boll rot. Seven varieties (FH-114, FH-113, BT-7, BT-212, SLH-BT-4, BT-212 and FH-941) were highly susceptible to bacterial seed and boll rot indicated by 2001–2300 AUDPC units. Biochemical tests identified bacterial isolates as Pantoea agglomerans. Different inoculation techniques were assessed for bacterial pathogenicity and symptoms of boll rot were only observed in needle punctured bolls. One, two and three weeks old bolls were mechanically inoculated by injecting bacterial suspension to evaluate the boll’s age impact on disease severity. Maximum severity was observed in two weeks old bolls. Red cotton bugs (Dysdercus cingulatus) were fed on artificially inoculated diseased bolls and then transferred on healthy bolls. Diseased symptoms were noticed on healthy cotton bolls. Bacterial colonies were recovered and red cotton bug was confirmed as the disease-transmitting vector.     

Bt抗虫杂交棉叶片营养元素含量变化及其与产量的关系

为了比较杂交棉不同品种、不同世代棉花叶片营养元素含量的差异,于2003、2004年以Bt抗虫杂交棉NZ62、H64、H16的杂交种一代、二代和三代为材料,在湖南长沙进行了研究。田间试验在湖南农业大学试验基地进行,采用随机区组试验,设4次重复,4行区,小区面积13．4m^2。分别于棉花盛花期、吐絮期两个时期,在每个试验小区连续取10株棉株的倒数第4叶和倒数第2叶,测定棉叶的全量N、P、K、B、Cu、Zn、Fe、和Mn的含量。研究结果表明：不同品种花铃期养分含量除P以外均存在显著差异,且变化趋势基本一致,均以NZ62含量最高,H16次之,H64最低,差异达显著水平（P〈5％）。吐絮期不同品种叶片的N、P、K和B含量均不存在显著差异,而Cu、Zn、Mn营养含量均存在显著差异,NZ62比H16和H64高;不同世代花铃期F1棉株叶片的N、K、Cu、Zn、Fe、Mn和B的含量显著高于F2、F3（P〈5％）,而F2、B的含量变化基本一致,F1代棉株对营养元素的吸收与F2、F3代棉株相比具有显著的杂种优势。F2、F3代优势衰退与其叶片营养元素含量减少相符合。但吐絮期不同世代之间没有显著差异。相关分析表明,花铃期棉花叶片含N量、含K量与子棉产量和皮棉产量有显著相关（P〈5％）,而吐絮期的这两个元素的含量与棉花产量无显著相关（P〉5％）。无论是花铃期还是吐絮期,B元素的含量与子棉、皮棉产量的相关均达到极显著水平（P〈1％）。     

棉铃发育期棉花源库活性对棉铃对位叶氮浓度的响应

采用大田试验,以3个铃期差异明显的棉花品种为材料,研究了不同施氮量形成的棉铃对位叶氮浓度对棉花花铃期纤维发育源库活性指标的影响。结果表明:在花后同一时期,棉铃对位叶可溶性糖、蔗糖含量和磷酸蔗糖合成酶活性以及棉纤维蔗糖含量和蔗糖合成酶活性等均随对位叶氮浓度的升高呈先升高后降低的变化趋势(45、52 DPA(花后天数Days post anthesis)的纤维蔗糖含量趋势相反),可用抛物线方程Y=ax2+bx+c拟合(P<0.01),通过拟合方程得到各指标所对应的最佳对位叶氮浓度。45 DPA(德夏棉1号38 DPA)前,花后同一时期各指标对应的最佳对位叶氮浓度差异较小,通过幂函数方程建立最佳叶氮浓度随花后天数的拟合方程,得到纤维发育期内源库活性各指标达到或接近最优状态时的适宜对位叶氮浓度的动态方程。本试验条件下,德夏棉1号、科棉1号和美棉33B的适宜对位叶氮浓度的拟合方程分别为N德1=7.2263DPA-0.276(R2=0.9805**)、N科1=7.23DPA-0.3026(R2=0.9861**)、N美33B=7.0997DPA-0.2814(R2=0.9807**)。     

Disease forecasting model for newly emerging bacterial seed and boll rot of cotton disease and its vector (Dysdercus cingulatus)

Bacterial seed and boll rot disease is a newly emerging threat to the cotton growers. Disease prediction model was devised to predict the disease progression impacted by the vector (Dysdercus cingulatus) and environmental variables (maximum air temperature, minimum air temperature, relative humidity and rainfall) on four varieties to minimise its losses and disease management cost. Impact of a-biotic environmental variables (maximum and minimum air temperature, relative humidity and rainfall) was assessed on bacterial seed and rot of cotton disease and its vector (D. cingulatus) on FH-941, FH-942, MNH-886 and FH-114 cotton varieties. Maximum red cotton bug population was assessed at 29–31 °C maximum temperature and at 15–17 °C minimum temperature. Disease severity was noticed maximum when maximum and minimum temperature was measured at 28–29 °C and 13–14.5 °C, respectively. Vector population was maximum when relative humidity and rainfall were 63–66% and 1.50–2.5 mm, respectively. Relative humidity at 66–68% and 0.5–1.5 mm rainfall favoured disease development. With increase in number of bugs, increase in disease severity was noted, maximum disease severity 45–48% noticed when 7–8 bugs were recorded. Red cotton bug (Dysdercus cingulatus) population prediction model was devised based on a-biotic factors (maximum and minimum air temperature, relative humidity and rainfall) on four cotton varieties. Disease forecasting model was developed based on biotic (D. cingulatus) and a-biotic factors. A close resemblance between observed and the predicted red cotton bugs and disease severity was seen.     

施氮方式对转基因棉花Bt蛋白含量及产量的影响

为研究氮肥运筹对棉花叶片、棉蕾和棉铃不同器官中Bt蛋白含量的影响,2009—2010年,以抗虫杂交棉中棉所72为试验材料,在大田条件下进行了不同基肥：花铃肥：盖顶肥施氮比例（分别为0:0.4:0.6、0.2:0.4:0.4、0.4:0.4:0.2、0.6:0.4:0）的试验。结果表明,施氮方式对棉花不同器官中Bt蛋白含量有明显影响。总体表现为随着氮肥前移,棉花幼嫩器官中Bt蛋白含量呈明显增加的趋势,而老熟器官中Bt蛋白含量呈明显降低的趋势。施氮方式对棉花幼嫩器官中Bt蛋白含量的影响比老熟器官明显,尤其是对幼嫩叶片Bt蛋白含量的影响大于幼小的棉蕾和棉铃器官。抗虫棉采用基肥：花铃肥：盖顶肥为0.4:0.4:0.2的施氮方式,总体能提高前中期棉花器官Bt蛋白的含量,有利于提高其抗虫性能;减少后期棉花器官Bt蛋白的含量,减轻对环境的压力;而且比其余3种施氮方式的籽棉产量和皮棉产量分别增加4.15%—11.24％、3.73%—12.01%。     

转基因抗虫抗除草剂棉花对棉铃虫抗性及对草甘膦耐受性研究

本论文研究了抗虫又抗除草剂棉花对草甘膦的耐受程度,比较了这一双抗性状的棉花与单抗虫棉的抗虫效果。结果表明,抗虫又抗除草剂棉花对草甘膦有较好的耐受性,四叶期喷施草甘膦后抗虫抗除草剂棉花可以安全生长,蕾期喷施草甘膦对棉花的开花率和结铃率有影响。抗虫又抗除草剂棉花和单抗虫棉对棉铃虫Helicoverpa armigera(Hübner)均具有较好的防治效果,苗期棉花叶片对棉铃虫防治效果最好,后期防治效果下降到49.2%和46.6%,吐絮期防治效果又上升到57.0%和53.1%。     

不同性状的棉花及其杂交F1代对棉铃虫的抗性

通过连续两年的田间试验,系统、全面地研究了棉花对棉铃虫(Helicoverpa armigera)的形态抗虫作用。研究结果表明：在小区试验条件下,以百株累积卵量、百株平均幼虫量及百株平均蕾铃受害量为考察指标,红叶可使危害减轻28.1%、31.45%和36.O%;鸡脚叶可使其下降19.0%、25.4%和28.4%;窄卷苞叶使其降低25.8%、32.9%和34.0%;光叶使各指标分别降低32.7%、35.1%和37.3%。高棉酚(油腺)抗性主要表现为对幼虫的抗生作用,使幼虫量下降了45.1%,使蕾铃受害量下降了51.4%。此外,研究还表明,两种抗虫性状的抗性效果有累加作用,鸡脚叶与红叶联合作用使三项指标分别下降达48.3%、55.2%和57.9%;红叶与窄卷苞叶则使危害减轻了52.7%、58.3%和60.6%;具有光叶与较密油腺两个抗虫性状的品种受害可减轻29.9%、54.9%和58.3%。将一些具有相对性状的品种进行杂交,其F₁有不同的抗虫表现。红叶、鸡脚叶和光叶品种与其对照杂交后的F1代因在不同程度上保持了抗虫性状故能表现出抗性,而窄卷苞叶品种与其对照杂交后F1代苞叶正常,则未能表现出抗性。因小区条件限制了无蜜腺抗性的表达,无蜜腺小区未表现出明显差异。     

Screening for High-Temperature Tolerant Cotton Cultivars by Testing In Vitro Pollen Germination, Pollen Tube Growth and Boll Retention

With radical global climate change and global warming, high temperature stress has become one of major factors exerting a major influence on crop production. In the cotton (Gossypium hirsutum L.)-growing areas of China, especially in the Yangtze River valley, unexpected periodic episodes of extreme heat stress usually occur in July and August, the peak time of cotton flowering and boll loading, resulting in lower boll set and lint yield. Breeding programs for screening high temperature-tolerant cotton germplasm and cultivars are urgent in order to stabilize yield in the current and future warmer weather conditions. In the present study, 14 cotton cultivars were quantified for in vitro pollen germination and pollen tube growth in response to temperatures ranging from 10 to 50 ℃ at 5 ℃ intervals. Different cotton genotypes varied in their in vitro pollen germination and pollen tube length responses to the different temperatures. Maximum pollen germination and pollen tube length ranged from 25.2% to 56.2% and from 414 to 682 μm, respectively.The average cardinal temperatures (Tmin, Topt, and Tmax) also varied among the 14 cultivars and were 11.8,27.3, and 42.7 ℃ for pollen germination and 11.8, 27.8, and 44.1 ℃ for maximum pollen tube length. Variations in boll retention and boll numbers per plant in field experiments were found for the 14 cotton cultivars and the boll retention and boll retained per plant on 20 August varied considerably in different years according to weather conditions. Boll retention on 20 August was highly correlated with maximum pollen germination (R2=0.84) and pollen tube length (R2=0.64). A screening method based on principle component analysis of the combination of pollen characteristics in an in vitro experiment and boll retention testing in the field environment was used in the present study and, as a result, the 14 cotton cultivars could be classified as tolerant, moderately tolerant, moderately susceptible and susceptible to high temperature.     

棉花衰老和熟相形成的生理生态与调控研究进展

衰老是棉株或其某些器官发生导致生命活动自然终止的败坏过程,是棉株生长发育过程的必然归宿,熟相则是棉株吐絮成熟期的表现,是衰老的表现形式和结果,有早衰、贪青晚熟和正常成熟之分.衰老和熟相都是基因型与环境互作的结果.本文总结评述了棉花衰老的生理生态和分子生物学方面的研究进展,提出了依靠选育稳发型棉花品种、合理使用植物生长调节剂并综合运用农艺栽培措施调控棉株生长发育和衰老,实现正常熟相,进而提高棉花产量和品质.     

Comparison of Resistance to Cotton Leaf Curl Disease (Multan/Burewala) in Gossypium hirsutum L. Varieties and Breeding Lines

Twenty‐four cotton varieties and advance breeding lines were evaluated for their resistance to cotton leaf curl disease (CLCuD; Multan/Burewala) under natural field and in glasshouse conditions for two consecutive years. Resistance was based on symptom expression and disease severity index. All the cotton genotypes exposed to the vector whitefly in the field and artificially infected by grafting manifested a high level of resistance against CLCuD (Multan) with the exception of genotype NIAB‐999 that was moderately resistant. All the test varieties/breeding lines were highly susceptible to CLCuD (Burewala) both in the field and the glasshouse. However, substantial differences were noted between genotypes for disease index under field conditions. Graft inoculation studies showed that all genotypes inoculated with CLCuD (Burewala) developed disease within 9–13 days whereas those graft‐inoculated with CLCuD (Multan) developed symptoms from 15 to 22 days after grafting. Severe reduction occurred in plant morphology, fibre and yield parameters of cotton variety NIAB‐111 following inoculation with CLCuD (Burewala) as compared with CLCuD (Multan).     

施氮量和种植密度对东北特早熟棉区棉铃生物量和氮素累积的影响

以辽棉19号和美棉33B为材料,研究了不同施氮量(0、240、480 kg ·hm-2)和不同种植密度(75000、97500、120000 plants·hm-2)对东北特早熟棉区棉花棉铃生物量和氮素累积特征的影响.结果表明： 棉花单铃、棉籽和纤维的生物量及其氮素累积随棉花生育进程的动态变化均符合“S”型曲线,种植密度和施氮量可以显著影响棉铃各部分生物量和氮素累积的动态特征,以及棉花产量与品质;在施氮量240 kg·hm-2和种植密度97500 plants·hm-2处理下,单铃、棉籽和纤维的生物量均达到最大,生物量和氮素累积的快速累积起始时间和终止时间较早但持续时间较短,生物量快速累积速率最大,生物量和氮素在铃壳中的分配系数最低,在棉籽和纤维中分配系数最高.     

Physiological Effects of Meloidogyne incognita Infection on Cotton Genotypes with Differing Levels of Resistance in the Greenhouse

Greenhouse tests were conducted to evaluate (i) the effect of Meloidogyne incognita infection in cotton on plant growth and physiology including the height-to-node ratio, chlorophyll content, dark-adapted quantum yield of photosystem II, and leaf area; and (ii) the extent to which moderate or high levels of resistance to M. incognita influenced these effects. Cultivars FiberMax 960 BR (susceptible to M. incognita) and Stoneville 5599 BR (moderately resistant) were tested together in three trials, and PD94042 (germplasm, susceptible) and 120R1B1 (breeding line genetically similar to PD94042, but highly resistant) were paired in two additional trials. Inoculation with M. incognita generally resulted in increases in root gall ratings and egg counts per gram of root compared with the noninoculated control, as well as reductions in plant dry weight, root weight, leaf area, boll number, and boll dry weight, thereby confirming that growth of our greenhouse-grown plants was reduced in the same ways that would be expected in field-grown plants. In all trials, M. incognita caused reductions in height-to-node ratios. Nematode infection consistently reduced the area under the height-to-node ratio curves for all genotypes, and these reductions were similar for resistant and susceptible genotypes (no significant genotype × inoculation interaction). Our study is the first to show that infection by M. incognita is associated with reduced chlorophyll content in cotton leaves, and the reduction in the resistant genotypes was similar to that in the susceptible genotypes (no interaction). The susceptible PD94042 tended to have increased leaf temperature compared with the genetically similar but highly resistant 120R1B1 (P < 0.08), likely attributable to increased water stress associated with M. incognita infection.     

花铃期土壤持续干旱对棉铃对位叶气体交换参数和叶绿素荧光特性的影响

在盆栽条件下,以杂交棉泗杂3号为材料,以花铃期正常灌水\[土壤相对含水量(SRWC)(75±5)%\]为对照,设花铃期SRWC(60±5)%和SRWC(45±5)%持续干旱50 d两个处理,研究棉铃对位叶气体交换参数和叶绿素荧光参数在持续干旱过程中的动态变化和响应机制.结果表明: SRWC (60±5)%处理0～21 d棉铃对位叶的净光合速率(P_n)、气孔导度(g_s)、胞间CO₂浓度(C_i)下降,气孔限制值(L_s)上升,叶绿素荧光参数无显著变化,P_n下降的主要原因是气孔限制;处理21～49 d棉铃对位叶P_n持续下降,C_i上升,L_s下降,同时最大光化学效率(F_v/F_m)、实际光化学效率(Φ_PSII)和光化学猝灭系数(q_P)显著降低,非光化学猝灭系数(NPQ)先升后降,P_n下降的主要原因是非气孔限制;此时叶片PSII系统受到损伤,光合机构及光合酶系统被破坏,同时成铃强度急剧下降,成铃数降低,导致产量下降.SRWC(45±5)%处理0～14 d棉铃对位叶P_n、g_s、C_i显著下降,L_s急剧上升,F_v/F_m、Φ_PSII、q_P无显著变化,P_n下降主要由气孔限制引起;处理14～49 d,棉铃对位叶P_n缓慢下降,C_i上升,L_s下降,F_v/F_m、Φ_PSII和q_P不断降低,而NPQ先升后降,表明P_n下降主要由非气孔限制引起,同时成铃强度急剧下降,成铃数减少,产量降低.本试验条件下,SRWC(60±5)%和SRWC(45±5)%处理下棉花生长的临界胁迫时间分别为21和14 d.     

Role of Pantoea agglomerans in opportunistic bacterial seed and boll rot of cotton (Gossypium hirsutum) grown in the field

AIMS: To investigate the aetiology of seed and boll rot of cotton grown in South Carolina (SC). METHODS AND RESULTS: Bacteria were isolated from diseased locules of cotton bolls collected in a field in SC, USA and tested for the ability to cause comparable disease symptoms in greenhouse grown cotton fruit. Spontaneously generated rifampicin-resistant (Rif(r)) mutants of the isolates were used in confirmatory pathogenicity tests. Resistance to the antibiotic was both stable and effective in differentiating between an inoculated Rif(r) strain, rifampicin-sensitive contaminants and/or endophytes. A series of inoculation methods was tested at various boll developmental stages and at different fruiting nodes on the plant. Field disease symptoms were reproduced by inoculating bolls at 2 weeks postanthesis with bacterial suspensions ranging from 10(3) to 10(6) CFU ml(-1). Pathogenic isolates were categorized as Pantoea agglomerans on the basis of phenotype testing, fatty acid profiling (similarity index = 0.94), and 16s ribosomal DNA sequence analysis (99% nucleotide identity). CONCLUSIONS: Pantoea agglomerans isolates from field-collected immature, diseased cotton caused comparable infection symptoms in greenhouse produced cotton fruit. SIGNIFICANCE AND IMPACT OF THE STUDY: In 1999, significant yield losses in SC cotton resulted from a previously unobserved seed and boll rot that has since been reported in other southeastern states. This study demonstrated a role of P. agglomerans in producing opportunistic bacterial seed and boll rot of cotton.