Rapid Screening of Cacao Genotypes for Field Resistance to Phytophthora palmivora Using Leaves, Twigs and Roots

Black pod, caused by Phytophthora spp. is one of the most important diseases of cacao occurring worldwide. Losses due to black pod caused by P. palmivora are still moderate in Côte d'Ivoire but P. megakarya causes high losses in Ghana and other Central African countries. Variation in field attack has been observed between cacao genotypes, but evaluation of pod losses is unsuitable for obtaining rapid progress in breeding. Results of inoculation tests using young detached leaves, twigs and roots, obtained from field and nursery plants, are presented here and compared to field resistance of similar genotypes observed over a 10-year period. Nine different Upper Amazon Forastero genotypes were tested together with progenies obtained by crossing these with the susceptible check IFC5 (Amelonado genotype). Rank correlations between the early screening tests and the level of field attack were positive and mostly significant (r=0.58–0.95). The coefficient of correlation was slightly higher for leaves (r=0.88) and roots (r=0.89) than for twigs (r=0.76). Also, resistance of the different plant organs was correlated (r=0.6–0.9). Resistance of the Upper Amazon parents was well correlated with the resistance of their cross progenies (r=0.7–0.9), suggesting that resistance is highly heritable. Resistance of leaves and twigs from the nursery was better correlated with field resistance than resistance of leaves and twigs from the field, which might result from more uniform growing conditions in the nursery. Inoculation of leaves appears the most suitable early screening method for black pod resistance. Application of this test in breeding more resistant cacao cultivars is discussed.     

园龄对可可苗期生长及其根际土壤环境的影响

可可是世界最重要的多年生经济作物之一,为了揭示不同园龄可可园土壤演变规律,为其可持续生产提供依据。以休耕地为对照(CK),研究低龄(4 a)、中龄(12 a)和老龄(32 a)3种不同园龄土壤培育可可苗期生长及其与根际土壤有机碳(SOC)含量、土壤酶活性和微生物区系等关系。结果表明,32 a园龄土壤培育可可苗生物量显著高于4 a、12 a和CK,分别增加了89.15%、29.24%和28.03%;32 a和12 a园龄处理根际SOC含量显著高于4 a园龄,老龄园有利于SOC的积累;随着园龄增加,脲酶活性增加,且与SOC含量极显著正相关,12 a和32 a园龄土壤脲酶活性分别比CK显著增加了44.36%和43.11%;12 a园龄处理根际土壤中可培养细菌数量减少,真菌数量则显著增加,12 a土壤可培养真菌数量分别是4 a、32 a和CK的3.04、1.46和3.23倍,园龄可能导致土壤微生物区系变化。综上所述,随着土壤园龄增加,可可苗生物量、根际SOC含量和脲酶活性均增加,根际土壤微生物区系发生变化,根际土壤真菌数量与SOC含量、脲酶活性极显著正相关,可可根际SOC含量是光合作用、植株生长、土壤酶活性和土壤微生物共同作用的结果。     

Spatiotemporal pattern analysis of Cacao swollen shoot virus in experimental plots in Togo

In order to quantitatively analyse the spatial pattern of cacao swollen shoot disease, particularly in cases of re‐emergence, three experimental plots were installed in a diseased area of cacao cv. Amelonado in Togo. After thorough cleaning and grubbing, the three plots were planted with less susceptible, hybrid plant material. Twenty years after replanting, a survey of healthy, diseased and dead trees was carried out during 2 consecutive years. Data were analysed using Ripley’s functions and join counts analysis. The re‐emergence of the disease occurred in patches or foci: following analyses with the two statistical methods, diseased trees and dead trees were found to be clearly aggregated on the three observed plots for the 2 years. The observed progress of the disease was not the same on the three plots and seemed dependent on the disease state of the first year: the higher the attack rate of the first year, the faster the disease progression. The use of less susceptible plants helped keep the land productive for 15 years. In conclusion, uprooting of the first infection focus can extend the life of cacao plots.     

The Trichoderma koningii aggregate species

The morphological concept of Trichoderma koningii is found to include several species that differ from each other in details of phenotype (including conidium morphology, growth rate) and biogeography. Phylogenetic analysis utilizing partial sequences of the translation-elongation factor 1 alpha (tef1), as well as fragments of actin and calmodulin genes, indicate that phenotypic characters typical of T. koningii evolved independently in three well-separated main lineages. Combined molecular and phenotype data lead to the development of a taxonomy with the recognition of twelve taxonomic species and one variety within the three lineages. These lineages include: (1) T. koningii and T. ovalisporum and the new species T. caribbaeum var. caribbaeum, T. caribbaeum var. aequatoriale, T. dorotheae, T. dingleyae, T. intricatum, T. koningiopsis, T. petersenii and T. taiwanense; (2) the new species T. rogersonii and T. austrokoningii, and (3) the new anamorph T. stilbohypoxyli.Trichoderma koningii s. str. is an uncommon species restricted to Europe and eastern North America; T. caribbaeum var. aequatoriale, T. koningiopsis, and T. ovalisporum were isolated as endophytes of trunks of Theobroma species in tropical America, and T. ovalisporum from the woody liana Banisteropsis caapi in Ecuador; T. koningiopsis is common in tropical America but was isolated also from natural substrata in East Africa, Europe and Canada, and from ascospores in eastern North America, and as an endophyte in Theobroma species; T. stilbohypoxyli, originally described as a parasite of Stilbohypoxylon species in Puerto Rico, is found to be more common in the tropics, besides an endophytic isolate from Fagus in U.K. The additional new species are known almost exclusively from their teleomorphs. Isolates of T. ovalisporum and T. koningiopsis may have biological control potential. A morphophenetic key and a set of tools for molecular species identification were developed.Taxonomic novelties:Trichoderma austrokoningii/Hypocrea austrokoningii Samuels & Druzhinina sp.nov., T. caribbaeum var caribbaeum/H. caribbaea Samuels& Schroers sp.nov., T. caribbaeum var. aequatoriale Samuels & H.C. Evans var.nov., T. dingleyae/H. dingleyae Samuels & Dodd sp.nov., T. dorotheae/H. dorotheae Samuels & Dodd sp.nov., T. intricatum/H. intricata Samuels & Dodd sp.nov., T. koningiopsis/H. koningiopsis Samuels, C. Suarez& H.C. Evans sp.nov., T. petersenii/H. petersenii Samuels, Dodd & Schroers sp.nov., T. rogersonii/H. rogersonii Samuels sp.nov., T. stilbohypoxyli Samuels & Schroers sp.nov., T. taiwanense/H. taiwanensis Samuels & M.L. Wu sp.nov.     

可可FAT基因家族进化及表达模式分析

在植物中,酰基-ACP硫酯酶（fatty acyl-ACP thioesterase, FAT）是调控脂肪酸合成的关键酶。为解析可可FAT基因家族成员的特点与功能,本研究从可可基因组中筛选鉴定出FAT基因家族的6个成员,分别命名为TcFATA、TcFATB1、TcFATB2、TcFATB3、TcFATB4、TcFATB5。6个基因外显子数目为6~7个,编码区（CDS）长度介于1128~1263 bp,预测蛋白分子量介于42.72~46.47 kDa,等电点介于6.57~9.10。进化分析结果表明可可FAT基因家族分成FATA与FATB亚群,FATA亚群包含1个可可TcFATA成员,FATB亚群包含5个可可TcFATBs成员。不同可可种子发育时期表达分析结果表明:TcFATA和TcFATB1伴随果实发育成熟,表达量呈下降趋势,TcFATA和TcFATB1在不同种质中表达量与油酸（C18:1）和棕榈酸（C16:0）比例呈正相关,表明其与脂肪酸组分比例调控有紧密关联。     

Validity of a leaf inoculation test to assess the resistance to Phytophthora megakarya in a cocoa (Theobroma cacao L.) diallel mating design

Cocoa pod rot, caused by different Phytophthora species, is rife in all production zones. Phytophthora spp. control is therefore a major challenge for world cocoa cultivation, and selection of resistant material is a priority research theme for many producing countries. Various genetic trials have been set up over many years in zones affected by the disease. In Cameroon, a 6x6 diallel mating design was set up to study heritability of the resistance trait. Field observations led to the classification of parents according to their ability to transmit resistance to their progenies. These field measurements were long and laborious, so a resistance test based on artificial leaf inoculations was developed. This article studies the correspondence between results obtained with the test and observations of rot rates in the field. There was a significant genetic correlation between pod rot rate in the field and leaf test data, so using this resistance test could shorten cocoa tree selection cycles. This revised version was published online in July 2006 with corrections to the Cover Date.     

Identification of a major QTL in cocoa (Theobroma cacao L.) associated with resistance to witches' broom disease

The witches’ broom disease caused by the fungus Crinipellis perniciosa is the main limiting factor for cocoa production in South America and the Caribbean. In Brazil, this disease affects almost all cocoa‐growing regions, causing serious economic, social and ecological damage. The aim of this study was to map genomic regions associated with resistance to C. perniciosa using an F₂ population derived from a cross between ‘Scavina‐6’(resistant) and ‘ICS‐1’(susceptible). The phenotypic index was determined as the average number of vegetative witches’ brooms per canopy area of each plant, the witches’ brooms were counted and eliminated during six field evaluations between May 1998 and August 1999. A total of 124 random amplified polymorphic DNA (RAPD) and 69 amplified fragment length polymorphism (AFLP) markers were mapped along 25 linkage groups covering 1713 cM of cocoa genome. After employing single factor and composite interval mapping analyses, a major quantitative trait loci (QTL) flanked by the marker AV14.940 was identified in the linkage group 11, explaining almost 35% of the resistance to witches’ broom. The present result suggests that this QTL acts as a major dominant component of resistance to this pathogen, with great potential for use in marker‐assisted selection procedures in cocoa breeding programmes.     

Competition effects in cocoa (Theobroma cacao L.) hybrid trials

A method developed in coffee tree breeding to assess family competitioneffects (partner effects) in comparative variety trials was applied to thecocoa tree (Theobroma cacao L.). The study was conducted in ahybrid comparative trial planted in French Guiana, involving twelve familiesof 50 trees in a totally randomized single-tree plot design, at a density of1,667 trees per hectare. The trial was thinned at 10 years, at a rate of twoout of four rows. Competition was studied with reference to juvenile andadult vegetative vigour, and to periodic and cumulative yields (number ofpods, potential weight, average weight of one pod, and the production:vigour ratio). At the end of the trial, after thirteen years of monitoring,competition effects were revealed which explained 8 to 10% of theresidual variance after removal of the hybrid and micro-environment effects.These effects, which were detected as early as 18 months, occurred earlierthan generally acknowledged. Under the trial conditions, the families couldbe classed as aggressive, stimulating or passive for their neighbours.Vegetative vigour (trunk cross-section) explained 34% of the competitioneffects (partner effects), which, with hindsight, vindicated the use of theproduction : vigour ratio as the main selection criterion in cocoa breeding.The partner effects noted on the production variables were never explainedby any production variable, hence non-aggressive high-yielding families canbe selected.     

Use of irradiated pollen as mentor pollen to induce self-fertilization of two self-incompatible Upper Amazon cacao clones

Summary Two self-incompatible Upper Amazon cacao clones, T85/799 and T79/501, were pollinated with compatible Amelonado pollen subjected to varying doses of gamma irradiation (10–100 Gy). The proportion of flat non-viable beans to fully formed, viable beans in the pods increased with an increase in dosage of gamma rays. At 60 Gy all the beans produced were flat and non-viable, beyond this dosage fruit set was zero. Pollinating the self-incompatible cacao clones with a 1 : 1 mixture of compatible mentor pollen irradiated at 60 Gy and normal self pollen produced a mixture of flat, non-viable beans and fully-formed viable beans. Similar experiments using irradiated pollen with a marker gene suggested that the fully-formed viable beans resulted from selfing. Increasing the proportion of the radiation-treated compatible pollen in the mixture increased the number of fully-formed beans. However, when compatible pollen which had been treated either at 80 Gy or with temperatures of 35° C, 40° C and 45° C for periods of five, ten and fifteen minutes in factorial combination were mixed with self pollen, no successful pollinations were achieved. Pollen viability tests indicated that, whilst pollen treated at 60 Gy were about 50% viable, those treated at either 80 Gy or with temperatures of 35–45° C were mostly not viable. This suggests that, to overcome the incompatibility in cacao, the tubes of the mentor pollen grains used should at least grow into the style. The possible causes for overcoming the self-incompatibility in cacao are discussed.