Inter Simple Sequence Repeat (ISSR) Analysis of Diploid Coffee Species and Cultivated Coffea arabica L. from Tanzania

Inter simple sequence repeat (ISSR) markers were used to evaluate levels of genetic similarity among Coffea arabica L. accessions from Tanzania and to estimate levels of genetic similarities in C. arabica and diploid coffee species. The six ISSR primers used generated a total of 82 fragments and the dissimilarity values ranged from 0.21 to 1. Mean dissimilarity values between provenances (0.56–0.85) were higher than within provenances (0.37–0.68). Cluster analysis based on Nei’s genetic distances showed C. arabica provenances grouping based on geographical origin. Two major clusters were formed that constituted of provenances from Kilimanjaro and Arusha in one sub-cluster; Tanga and Morogoro in the other; the second cluster had Mbeya provenances and diploid species, respectively. The implication is that Mbeya provenances are different from the rest of Tanzanian C. arabica. A principal coordinate analysis (PCA), whose first three coordinates explained 43% of the variation, showed similar groupings as in the cluster analysis. A separate cluster analysis of diploid species showed a distinct separation of the three species used. ISSR data gave results similar to previous findings from random amplified polymorphic DNA(RAPD) analysis. The results also confirm the limited diversity present in cultivated C. arabica in Tanzania     

土壤干旱对小粒种咖啡有关生理参数的影响

对塑料袋一年生咖啡苗定期停止淋水，使土壤遭受不断加剧的干旱胁迫，以检测水分供应对咖啡生理状况的影响。试验结果表明：轻度水分胁迫，如１ｄ不淋水，即导致咖啡光合速率、蒸腾速率、可溶性蛋白质和叶绿素、类胡萝卜素含量以及含水量、气孔开张率、水势等生理参数开始下降；而过氧化物酶活性、脯氨酸和丙二醛含量以及细胞透性等生理参数开始上升，且随土壤水分胁迫的逐步加剧，上述各生理参数呈渐进性恶变。光合速率对土壤水分的反应属敏感参数之一，轻度干旱即使光合速率明显下降。高温间歇干旱期，常规管理的田间含水量能使咖啡光合速率受到抑制，灌水可促使光合速率提高。     

Genomic variability and plasticity of Pseudomonas causing coffee leaf spots in Minas Gerais state,Brazil

Foliar spots caused by Pseudomonas coronafaciens pv. garcae (Pcg), Pseudomonas amygdali pv. tabaci (Pat) and Pseudomonas cichorii (Pch) are major bacterial diseases that can reduce coffee production. However, little is known about the genetic diversity and molecular mechanisms underlying the pathogenicity to coffee plants of these bacteria. In this study, genome sequences of Pcg, Pat and Pch strains isolated from coffee plants in Minas Gerais state, Brazil, were used to assess their variability and plasticity, and compare their type III secretion system (T3SS) and apoplastic effector repertoires as well as tabtoxin biosynthetic/detoxification genes. Genomic diversity was found for all three phytopathogens, among which Pch possesses the highest number of exclusive proteins. The Pcg genome is the most stable whereas that of Pch is the most plastic, which is related to their host ranges. When compared with those of Pseudomonas syringae pv. tomato DC3000, hrp/hrc gene sequences are more conserved in Pcg and Pat than in Pch, which also possesses the smallest T3SS and the largest apoplastic effector repertoires. The only T3SS effector family common to all three pathogens is AvrE, suggesting that, as for other plant–Pseudomonas interactions, it may play a crucial role for pathogenicity towards coffee plants. Apoplastic proteins associated with maintaining the redox balance and degrading proteins/peptides not previously described as important in plant–bacteria interactions were found. Gain/loss of the tabtoxin biosynthetic cluster with retention of the antitoxin gene was observed, indicating that tabtoxin production is not a limiting factor for the occurrence of mixed infections.     

咖啡黑小蠹的发生规律及药剂防治研究

咖啡黑小蠢是近年在兴隆华侨农场严重危害中粒种咖啡的重要害虫。据1989年对4—5年生咖啡树调查,植株受害率100％,枝条受害率29.7—82.5％,虫害枯枝率7.65—17.8％。该虫以雌成虫钻蛀咖啡枝条为害,幼虫和成虫取食蛀道壁上的真菌菌丝。田间种群数量通常在3月上旬开始剧增,3月中下旬为高峰期,7月至10月田间虫口极少,11月以后逐渐有虫口及虫枯枝出现。温度是影响虫口波动的主要因素。2.5％溴氰菊酶、25％杀虫双、40％乐菊酯各兑水1000倍对成虫直接触杀效果显著,死亡率均为100％。48％乐斯本、40％氧化乐果400—800倍液等12种杀虫剂田间喷雾对咖啡枝条蛀洞内的成虫、蛹、幼虫防效均不明显。     

Kinetics of Zinc Uptake and Anatomy of Roots and Leaves of Coffee Trees as Affected by Zinc Nutrition

Abstract

The influence of silicon (Si) (2.5 mM), sodium chloride (NaCl) (100 mM), and Si (2.5 mM) + NaCl (97.5 mM) supply on chlorophyll content, chlorophyll fluorescence, the concentration of malondialdehyde (MDA), H₂O₂ level, and activities of superoxide dismutase (SOD; E.C.1.15.1.1.), ascorbate peroxidase (APx; E.C.1.11.1.11.), catalase (CAT; E.C.1.11.1.6.), guaiacol peroxidase (G-POD; E.C.1.11.1.7.) enzymes, and protein content were studied in tomato (Lycopersicon esculentum Mill c.v.) leaves over 10-day and 27-day periods. The results indicated that silicon partially offset the negative impacts of NaCl stress with increased the tolerance of tomato plants to NaCl salinity by raising SOD and CAT activities, chlorophyll content, and photochemical efficiency of PSII. Salt stress decreased SOD and CAT activities and soluble protein content in the leaves. However, addition of silicon to the nutrient solution enhanced SOD and CAT activities and protein content in tomato leaves under salt stress. In contrast, salt stress slightly promoted APx activity and considerably increased H₂O₂ level and MDA concentration and Si addition slightly decreased APx activity and significantly reduced H₂O₂ level and MDA concentration in the leaves of salt-treated plants. G-POD activity was slightly decreased by addition of salt and Si. Enhanced activities of SOD and CAT by Si addition may protect the plant tissues from oxidative damage induced by salt, thus mitigating salt toxicity and improving the growth of tomato plants. These results confirm that the scavenging system forms the primary defense line in protecting oxidative damage under stress in crop plants.     

INFLUENCE OF NITROGEN FERTILIZATION ON NICKEL ACCUMULATION AND CHEMICAL COMPOSITION OF COFFEE PLANTS DURING FRUIT DEVELOPMENT

Nutritional and physiological significance of micronutrients in coffee plants, especially with regard to nickel (Ni) is still unknown. The dynamics of nitrogen (N), phosphorus (P), potassium (K) and Ni accumulation in coffee fruits, as well as their relationships with total soluble protein, amino acids, reducing sugars, and starch content during coffee fruit development (green, ripe, and dry fruits), were investigated. Coffee trees received three N fertilizer rates (0, 150, and 300 kg of N ha^?1) as ammonium sulfate split into three applications per year. Nitrogen fertilization increased reducing sugars and starch concentrations in ripe fruits. In contrast, green fruits showed the highest amino acid and Ni concentrations. Fruit Ni concentration decreased in both green and ripe fruits as N rates increased; thus, indicating the possibility of either a N-associated dilution effect on Ni concentration or that Ni uptake by roots and/or transport to developing fruit was limiting. Plant nutritional status and fruit development stage influenced the coffee grain chemical composition. Furthermore, the variation in reducing sugars and starch content was more closely linked to the stage of fruit development than to N supply. A supposed relationship among the decreased of caffeine, starch, amino acids, and proteins with Ni content during green fruit development suggests a fundamental role for Ni in coffee fruit ripening. The interaction between N and Ni metabolism during fruit ripening might influence the chemical parameters involved in the coffee grain quality. This is the first report documenting changes in Ni concentrations of coffee fruit as a function of N fertilization rates and the development stage, but further research is needed to better understand the significance of N-Ni interaction in developing coffee fruit.     

Nutritional standards and nutritional diagnosis of the Conilon coffee plants in phenological growth stages

The objective of this study was to establish a nutrient sufficiency range and DRIS standards of Conilon coffee grown in southern Bahia, Brazil for the pre-flowering and grain-filling stages and to perform a nutritional diagnosis of these plants using different methods. The collections were performed in 24 Conilon coffee farm fields to establish the leaf nutritional standards, and 278 samples were collected to the nutritional diagnosis. The nutritional diagnosis was performed by using the sufficiency range criterion and the DRIS method. The nutrient sufficiency range and the DRIS standards must be specific to each phenological stage of the Conilon coffee plants. The leaf concentrations of calcium (Ca), sulfur (S), and boron (B) were higher in the pre-flowering stage; and the leaf concentrations of nitrogen (N), potassium (K), copper (Cu), and zinc (Zn) were higher in the grain-filling stage.     

Distribution analyses of Meloidogyne spp. and Pratylenchus coffeae sensu lato in coffee plots in Costa Rica and Guatemala

The distribution of the plant-parasitic nematodes Pratylenchus coffeae sensu lato and Meloidogyne spp. were studied in two plots, one in Guatemala ( P. coffeae and M. paranaensis ) and the other in Costa Rica ( P. coffeae and M. exigua ). The quantity of nematodes per g fresh weight root were counted for each coffee tree sampled. The distributions were aggregated, and generally fitted well to negative binomial distributions. Population aggregation was greater when a smaller number of nematodes were involved, suggesting that initial colonization develops in foci. Analyses of the relationships between population levels of the species suggested that there was competition between Pratylenchus coffeae and Meloidogyne spp. This competition was expressed differently depending on the relative population density of the different species.     

Identification of a major gene (Mex-1) from Coffea canephora conferring resistance to Meloidogyne exigua in Coffea arabica

Among the most damaging root-knot nematode species, Meloidogyne exigua is especially common in Latin America and constitutes a major agronomic constraint in all major coffee-growing ( Coffea arabica ) areas. Growing nematode-resistant coffee represents the most promising option for control of the pest. The present study aimed to determine the mode of inheritance of the M. exigua resistance transferred into C. arabica from a related species, Coffea canephora , and to identify associated molecular markers. Segregation data analysis of F ₂ progeny derived from a cross between the resistant introgression line T5296 and the susceptible accession Et6 showed that the resistance to M. exigua is controlled by a simply inherited major gene (designated the Mex -1 locus). The gall index distribution exhibited by the F ₂ individuals suggested incomplete dominant expression. Fourteen AFLP markers were found associated with the resistance to M. exigua and a localized genetic map of the chromosome segment carrying Mex -1 was constructed. Furthermore, the association of the identified AFLP markers with Mex -1 was confirmed by analysis of a set of genotypes involving 28 introgression Arabica lines either resistant or susceptible to M. exigua in field conditions. These results represent an important starting point to enhance backcross breeding programmes and to perform an early selection of resistant seedlings.     

手持振动梳刷式小粒咖啡采摘装置设计与试验

针对云南丘陵山区小粒咖啡人工采收成本高、大型机械采收困难等问题,设计了手持振动梳刷式小粒咖啡采摘装置。通过建立果树-机械收获动力学模型和多体动力学仿真,得到振幅的稳态响应表达式,并确定了振动部件和梳刷部件的结构参数。利用ADAMS建立了果树-机械刚柔耦合模型,采用广义力与传感器函数控制的方法进行单因素仿真试验,分析了频率、梳齿间距和偏心块夹角对小粒咖啡收获效果的影响规律。以频率、梳齿间距和偏心块夹角为试验因素,采净率、采青率和损伤率为试验指标,进行二次回归正交旋转组合试验,结果表明：对采净率和采青率的影响因素重要性依次为频率、梳齿间距、偏心块夹角;对损伤率影响因素重要性依次为梳齿间距、频率和偏心块夹角。当频率为26Hz、梳齿间距为32mm、偏心块夹角为22.5°时,采摘机工作性能最佳,经试验验证,此时采净率为91.35％,采青率为8.46％,损伤率为4.15％,满足小粒咖啡收获技术要求。