Genetic Analysis of Yield and Yield Components Based on the Three Controlled Hybrid Populations in the Robusta Coffee （Coffea canephora Pierre）

Genetic analysis of yield and yield components were carried out on three parental clones of Coffea canephora Pierre and their controlled hybrid progenies. There is a possibility to estimate coffee yield capabilities in simply via the easier measuring traits of yield components since significant correlation between them were detected. We found that among parental show very few significantly differences of all observed characters and even not found between the overall mean of the descendants and their donor parents, whereas some particular descendants show higher values rather than their donor parent in one or more characters. Increased value between the best parent to the best descendent were detected varies from zero to 102%. This situation suggests the difficulty to find out the optimum yield in a single plant by accumulating all of yield components that actually can increase 143% to 278% from actual to potential yields. In addition, observed characters among population were also actually found highly varies from 159% in out-turn in population B to 28,333% in number of productive branch among population A. Low heritability （h2） that generally found in quantitative traits due to polygenic control was present in our studies in almost all characters observed. Therefore, selection for actual high yielding through conventional breeding will not efficient, and more convincing the power of selection using molecular markers to assist for cumulate additive effects in a single genotypes that resulted of high yielding in the future.

Mapping of Quantitative Trait Loci （QTLs） Controlling Cherry and Green Bean Characters in the Robusta Coffee （Coffea canephora Pierre）

In coffee breeding practice, two quantitative traits, namely cherry and green bean characters are the important phenotypic selection index. The synchronous of cherry maturation, size, weight and shape of green bean are desirable traits for the future breeding. In order to increase the breeding efficiency, a set of quantitative trait loci （QTLs） analysis controlling these traits was carried out. The QTL analysis was performanced in cross pollinated population of Coffea canephora using single nucleotide polymorphisms （SNPs） and simple sequence repeats （SSRs） markers. Totally 12 QTLs determined desirable cherry and green bean traits were identified on 9 linkage groups （LG）, where these QTLs mapped forming clusters in 11 different chromosomal regions. The desirable traits which can be detected by their QTLs are synchronous of cherry maturation （LG B and LG G）, cherry size （LG A, LG F and LG G）, bean shape （LG B, LG D and LG J）, bean weight （LG H）, bean size （LG A and LG E） and cherry-bean size （LG K） The gene action of these QTLs was dominance or an interaction of alleles （epistasis between alleles at the studied locus） effect. The yielded linkage map and the detected QTL can provide a tool for further genetic analysis of two traits and be potential for maker-assisted selection in C. canephora breeding.

Identification of Quantitative Trait Loci （QTLs） Determining Flowering in the Robusta Coffee （Coffea canephora Pierre）

The quantitative trait loci （QTLs） analysis for flower traits were performed based on three crossing of Coffea canephora in Indonesia. This study successfully identified 19 QTLs related to flowering. These QTLs consisted of 1） three QTLs associated with green candle flowering time located at linkage groups of A and D; 2） one QTL associated to white candle flowering time located at linkage groups of B; 3） two QTLs associated to blossom flowering time located at linkage groups of B; 4） two QTLs associated to number of flowers located at linkage groups of G and H; 5） five QTLs associated to score of flowers quality located at linkage groups of A, C and D; 6） five QTLs associated to number of star flowers located at linkage groups of D, E, F and G. Due to some QTLs are overlapping on the same linkage group, thus, these 19 QTLs for flower traits might be considered as 15 different ones which distributed on 14 regions of the eight linkage groups. Although pleiotropic genes might be the most likely explanation for the collocation of these QTLs, the present data are not sufficient to distinguish between a pleiotropic gene and a cluster of different loci controlling several traits. These results can be a first step for molecular breeding on Robusta coffee mainly in order to improve potential suitable characteristics such as number of flowers and number of normal flowers as well as to reduce adverse characteristics such as number of star flowers.

Growth and Quality of Clonal Plantlets of Conilon Coffee(Coffea canephora Pierre ex A.Froehner)Influenced by Types of Cuttings

To achieve high crop yieldin agriculture, the production of plantlets of superior quality is one of thebasic prerequisites. For conilon coffee, the current recommendation in theproduction of clonal plantlets through vegetative propagation is to usediagonal cuttings, in bevel form. However, there are indications that othertypes of cutting have been successfully used to produce plantlets. In thiscontext, this experiment was conducted with the objective of studying thedevelopment of plantlets of conilon coffee obtained by different types ofcuttings. The experiment was conducted in a greenhouse. The cuttings wereobtained from the middle portion of orthotropic stems of standardized plants.The experiment followed a split plot in time design, with three types ofcuttings (bevel, sharp bevel and straight) and two times of evaluation, withfour repetitions. At 30 and 120 days after planting, plantlets were collectedand evaluated. The results show the increase occurred in the variables due tothe straight cut when compared to the others, except for stem diameter.Emphasizing in this context, the straight cut promoted an increase in seedlingheight, production of biomass and in the Dickson’s quality index when comparedto cuttings in bevel and sharp bevel.

Cellular Growth Dynamics Affects Allelopathic Activity in Coffee Cell Culture

Cellular growth dynamics and allelopathic activity in coffee cell cultures were examined as follows: First, we compared allelopathic activity of seven woody plant calli, Coffea canephora, Derris indica, Ficus carica L., Juniperus conferta, Prunus persica, Punica granatum, and Sonneratia ovata, using a modified “sandwich method bioassay” and found that coffee callus showed the strongest growth inhibition to lettuce seedling nearly 90% of hypocotyl and 96% of root. This coffee callus actively proliferated, with a 21-fold increase during five weeks of subculture, with a growth curve comprising two typical phases: a lag phase of 0 - 2 weeks of culture and an exponential phase of 3 - 5 weeks of culture. Allelopathic activity varied depending on the growth phase of the coffee callus. The strongest allelopathic activity was detected in 1 - 2-week-old callus showing nearly 100% inhibitory effect on lettuce seedling growth. As the allelopathic activity of coffee calli is extremely high, beyond the natural level in coffee leaves and green beans, we focused on analyzing the allelopathic activity of its aqueous extracts using high-performance liquid chromatography. Several prominent peaks, including two reference alkaloids, theobromine and caffeine, which are known allelochemicals in coffee plants, and three distinct unknown peaks were identified at 270 nm in coffee calli during the lag phase (1 - 2 weeks of culture). The higher value of the total phenolic content in the lag phase also suggested a key biosynthetic pathway in relation to the allelopathic activity of coffee callus will be activated in the lag phase.

单作和间作对槟榔和咖啡生长、根系形态及养分利用的影响

本研究设置了槟榔和咖啡单作及间作盆栽试验,对比分析了单作和间作种植形式下槟榔和咖啡生长量、生物量、根系形态、土壤和植株养分含量差异。结果表明:与单作相比,间作抑制了咖啡株高的生长,显著减少了咖啡的茎鲜重、干重,促进了槟榔株高、茎粗的生长,槟榔茎鲜重、干重显著增加;从槟榔咖啡根系空间分布可见,咖啡根系在盆栽土层内比槟榔根系分布范围宽,其根系延伸到槟榔根系生态位,能使该生态位槟榔根长密度增加,槟榔根系主要分布在本株根系空间范围内;间作显著提高了土壤速效磷、速效钾的含量;咖啡对氮、钾肥的吸收量大于槟榔,槟榔对钙、锌的吸收量大于咖啡,两者间养分吸收的差异,减少了作物间养分的竞争;间作咖啡氮平衡指数和叶绿素含量大于单作,间作槟榔叶片氮、磷,茎干氮、磷、钾,根氮、磷、钾累积量均大于单作。综上,槟榔间作咖啡根系互作具有对土壤养分资源利用的互馈效应。

干旱胁迫对中粒种咖啡幼苗膜脂过氧化、抗氧化酶活性和渗透调节物质含量的影响

以苗龄约8个月的中粒种咖啡袋育苗为材料．测定3种干旱胁迫强度下咖啡幼苗叶片的生理指标变化．旨在了解其对干旱胁迫的生理响应。结果表明：在轻度干旱胁迫下（土壤相对含水量为55％～60％）．与对照相比，中粒种咖啡幼苗叶片的可溶性蛋白、可溶性糖、脯氨酸含量、超氧化物歧化酶（SOD）、过氧化物酶（POD）、过氧化氢酶（CAT）活性极显著增加．而丙二醛（MDA）含量和相对电导率没有显著变化。但在中度干旱或重度干旱胁迫下（土壤相对含水量为40％．45％或25％～30％），可溶性蛋白含量、SOD、POD、CAT活性显著或极显著减少，而可溶性糖、脯氨酸的积累减缓，MDA含量和相对电导率迅速上升。在适度干旱胁迫下，中粒种咖啡幼苗可通过提高抗氧化酶活性和渗透调节能力来增强其对干旱胁迫的耐性。

利用SSR分子标记快速鉴别中粒种与小粒种咖啡

为快速、准确地鉴别中粒种与小粒种咖啡,以不同种源的中粒种和小粒种咖啡为试验材料,对514对SSR引物进行中、小粒种咖啡种间特异性引物筛选,最终筛选到1对特异性引物,该引物在小粒种咖啡样本中扩增出90 bp和110 bp双带产物,而在中粒种咖啡样本中只扩增出90 bp或100 bp的单带产物,根据产物条带的数目即可对中粒种和小粒种咖啡进行鉴别。经随机抽取中、小粒种咖啡样本进行种类鉴别准确性验证,验证结果与实际结果一致。

中粒种咖啡小孢子染色体加倍方法的研究

本文报道在中粒种咖啡(C.Canephora)花粉母细胞减数分裂时期,应用骤变低温和秋水仙素直接处理花器官,诱导形成二倍性花粉粒的方法。研究结果表明:(1)在花粉母细胞减数分裂时期用骤变低温处理可获得大量二倍性花粉粒。最佳处理时期为终变期——中期Ⅰ和后期Ⅰ;最适处理温度为8—10℃,对不同的无性系,处理效果有所不同。(2)应用1%浓度的秋水仙素滴液法直接处理花蕾可获得7.23—9.20%二倍性花粉。

咖啡种间嫁接与不嫁接植株主要农艺和品质性状比较

为探明种间嫁接生产应用可行性,以解决连作咖啡园再植植株长势弱、死亡率高的问题,布置了以未嫁接的中粒种咖啡热研3号实生后代为对照,查理种16号为砧木、中粒种咖啡热研3号实生后代为接穗的种间嫁接大田试验。经过5 a的田间性状观测,结果表明:1年生种间嫁接植株株高、茎粗、冠幅、最长一级分枝长度大于未嫁接植株,但差异未达显著,种间嫁接植株一级分枝对数、最长一级分枝节数显著大于未嫁接植株;2015年单节果实数未嫁接植株显著高于嫁接植株,其余年份(2014年、2016年)差异不显著;干鲜比、百粒重、单株鲜果产量两者间差异不显著。虽然嫁接植株2015年单节果实数显著低于未嫁接植株,但种间嫁接促进了中粒种咖啡的生长,增加了单株结果节数,因而嫁接植株单株鲜果产量未显著减少。综上,嫁接不影响中粒种单株鲜果产量和果实的品质性状,该组合的种间嫁接在生产中推广应用是可行的。

施用咖啡果皮对咖啡幼苗生长及土壤理化性状的影响

通过盆栽试验研究施用不同量的咖啡果皮对咖啡幼苗生长及土壤理化性状的影响。结果表明:施用5%(果皮与土壤质量比)以上咖啡果皮的咖啡幼苗地上部、根系干重,总根长,根系直径及根系总体积显著低于对照处理;随咖啡果皮施用量的增加,咖啡幼苗长势显著变差;施用5%以上的咖啡果皮可显著增加土壤碱解氮、速效磷、速效钾养分含量和土壤脲酶、酸性磷酸酶活性,但土壤p H显著下降。综上所述,施用5%以上直接堆沤腐熟的咖啡果皮会抑制咖啡幼苗生长,降低土壤p H,但可显著提高土壤养分含量和土壤脲酶、酸性磷酸酶活性,增加土壤养分有效性。

咖啡叶片DNA提取方法的比较研究

以咖啡成熟叶片为材料,进行多种DNA提取方法比较研究,并通过琼脂糖凝胶电泳和紫外分光光度计检测所提的DNA样品。结果表明,CTAB改良Ⅰ和CTAB改良Ⅱ均可用于咖啡叶片的DNA提取,尤其是CTAB改良Ⅱ法,提取的DNA产量较CTAB改良Ⅰ高,能满足后续分子生物学研究的要求。

中粒种咖啡光合特性的研究

自然状态下中粒种咖啡树冠外围向阳叶的光合速率及叶绿素 a/b 值均较树冠内部荫蔽叶大,但叶绿素含量和相等叶面积叶绿素的光谱吸收量却较小。与自然状态相同,人工遮荫叶的光合速率和叶绿素 a/b 值均较不遮荫的同龄叶小,但叶绿素含量和相等叶面积叶绿素的光谱吸收却较大。在低光照强度时,向阳叶和荫敝叶的光合速率部随光照强度的增加而提高,但光照强度超过一定值时,光合速率下降,即发生光抑制。发生光抑制的光照强度,向阳叶大于荫蔽叶。向阳叶光合作用的光饱和点和 DCPIP 光还原活性最大时的光照强度出都大于荫敝叶。呼吸作用和代表光呼吸的乙醇酸氧化酶活性也以树冠外围的向阳叶为大。中粒种咖啡的光合速率对低温敏感,在3万勒克司光照强度和高浓度 CO_2 条件下,光合作用的低温界限约在10℃。但温度高达35℃时,光合速率仍未见下降。

种间嫁接对中粒种咖啡品质的影响

为探明种间嫁接对中粒种咖啡品质的影响,采用大粒种(查理种)16号为砧木,中粒种热研3号为接穗进行种间嫁接大田试验,以未嫁接中粒种为对照。对嫁接与未嫁接的咖啡豆样品进行物理特性、化学组分含量和杯品品质测定和分析。结果表明:嫁接后大部分咖啡米粒径分布在>7.0 mm和>6.5 mm范围内,而未嫁接大部分咖啡米粒径分布在>6.0 mm和>5.0 mm范围内,嫁接显著增大了咖啡米粒度;咖啡豆百粒重和百粒密度两者间差异不显著;嫁接咖啡豆蛋白质、脂肪、咖啡因、绿原酸、葫芦巴碱含量分别为13.36%、5.40%、1.62%、5.12%、0.79%稍小于未嫁接,蔗糖含量8.19%,稍高于未嫁接,两者间差异未达显著;嫁接杯品为81.00分,未嫁接为82.25分,两者间差异不大,均达到≥80分的精品级别。可见,种间嫁接未对咖啡豆的饱满度、化学组分含量产生影响,咖啡杯品品质较好,该种间嫁接组合可推广生产应用。

咖啡中粒种和大粒种的核型研究

采用去壁、低渗、火焰干燥和Ｇｉｅｍｓａ染色制片法对中粒种咖啡（Ｃｏｆｆｅａｃａｎｅｐｈｏｒａ）和大粒种咖啡（Ｃ．ｌｉｂｅｒｉｃａ）的核型进行了分析，结果表明，中粒种咖啡的核型为２ｎ＝２２＝１０ｍ＋１２ｓｍ（２ＳＡＴ），大粒种的核型为２ｎ＝２２＝１２ｍ＋１０ｓｍ（２ＳＡＴ），根据Ｓｔｅｂｂｉｎｓ的核型分类标准，均属于２Ｂ类型。

18个中粒种咖啡无性系初级系比试验

进行了18个中粒种咖啡无性系的初级系比试验。结果表明:其中1个无性系综合性状表现较好,产量高于对照(已鉴定高产无性系“24-2”),3a平均鲜果年产量5.22kg/株;另外7个无性系枝节生长密集,树冠开张,株结果节数多,果丛大,产量较高,具有一定的利用潜力。

胡椒和咖啡2种中间性种子的萌发特性研究

研究了胡椒(Pipernigrum Linnaeus)和中粒种咖啡(Coffea canephora Pierre ex Froehner)种子在常温自然贮藏条件的萌发特性。结果表明,刚采收的胡椒种子和中粒种咖啡种子的含水量、发芽率、活力指数分别为29.32%和46.11%、88%和92.0%、1.46和5.57。自然贮藏20d,干燥脱水对胡椒种子的伤害较轻微,种子发芽率和活力指数仍高达82.7%和1.03;之后,由于严重脱水,种子发芽率和活力指数迅速下降;自然贮藏60d,种子含水量下降到10.91%时,完全丧失发芽力。自然贮藏30d,干燥脱水对中粒种咖啡种子的伤害较轻微,种子发芽率和活力指数仍高达80.5%和4.10;之后由于严重脱水,种子发芽率和活力指数急剧下降;自然贮藏90d,种子含水量下降到9.81%时,种子几乎完全丧失发芽力。由此认为,胡椒和中粒种咖啡是一种中间性种子,过度脱水使质膜结构和功能严重受损,最终导致种子活力的丧失。因此,在播种时,采用常温自然贮藏不超过20d的胡椒种子及贮藏不超过30d的中粒种咖啡种子为佳。

15个中粒种咖啡无性系的主要农艺性状观测

对15个中粒种咖啡无性系的主要农艺性状进行观测和综合评价,筛选出24-2号、兴28、兴29、26号、6号、24- 1号等产量高、农艺性状优良的无性系,建议生产上推广种植。

Coffee Agroforestry with Some Timber Shade Trees： Study on Carbon Stock, Mineral Cycle, and Yield

Coffee cultivation by using shade trees is simple of agroforestry, this system could get better ecosystem service and sustainable agricultural. The aims of this research are to study the possibility of some species of industrial woods as shade trees of Coffea canephora. The research was conducted in Jember, Indonesia （45 m asl., D rainfall type according to Schmidt and Ferguson）, and arranged in split plot design. The main plots were （A） coffee-T, grandis （3 m × 2.5 m ×12 m）, （B） coffee-P, falcataria single row （2.5 m ×6 m）, （C） coffee-P, falcataria double rows （3 m× 2.5 m × 12 m）, （D） coffee-P, falcataria vat. Solomon （3 m× 2.5 m × 12 m）, （E） coffee-M, azedarach （3 m ×5 m ×22.5 m）, （F） coffee-H, macrophyllus （3 m ×5 m ×12.5 m）, and （G） coffee-Leucaena sp. （3 m × 2.5 m） as control. The sub plots were coffee clones, i.e., BP 534, BP 409, BP 936, dan BP 939. Among those timber trees, Leucaena was planted as the alternative shade trees. The result showed that in comparison with control, all of coffee agroforestry system improved carbon sequestration. Total C-stock on （B） was highest, i.e., 1,007 percent to control while the lowest one was （A） 317.44% to control. During one year observation, litter weight of H. macrophyllus was heaviest followed by T. grandis. The lightest litter was obtained from M. azedarach. Based on its mineral contents, litters of T. grandis potentially supplied back nutrients that equaled to total Urea, SP-36, KC1, Dolomite, and Kieserite as much as 574.14 g; P. falcataria 287.57 g, P. falcataria var. Solomon 453.59 g, M. azedarach 450.84 g, H. macrophyllus 877.56 g, and Leucaena 445.12 g per tree per year. Because of heavily fallen leaves of M. azedarach during dry season and conversely too dense shading of H. macrophyllus, bean yield at 4 and 5 years old by using both species were consistently lower than that under T. grandis, P. falcataria and control. At those ages, effect of clone on cherry yield was still not consistent but there was a tendency that BP 939 was most productive, while BP 534 was the less. Its outturn was not influenced by agroforestry system but by clones. The agroforestry pattern influence physical bean characters, more dense of shading, more single bean and empty bean. That bean abnormality also genetically, on BP 939 percentage of round and empty bean was highest while on BP 936 was lowest. It was concluded that coffee agroforestry improve ecology service, but M. azedarach and H. macrophyllus were not appropriate to be used as coffee shade trees. P. falcataria is recommended as an alternative shade tree beside Leucaena sp.

Adaptation to Long-Term Rainfall Variability for Robusta Coffee Cultivation in Brazilian Southeast

Coffee is one of the world most traded agricultural commodities. Currently, a lot of attention has been on Robusta coffee (Coffea canephora Pierre ex A. Froehner) because it seems to evidence a greater tolerance to extreme climatic events than Arabica coffee (C. arabica L.). Despite this, only a few works have been developed aimed at discriminating the climatic vulnerability in regions which prioritize robust coffee production. The aim of this work was to analyze historical climatic variables in space and time for the characterization of climatic vulnerability of micro-regions, in search of mitigation and adaptation, which might support the improvement of production systems of C. canephora coffee trees. The case study was carried out for one of the largest production regions of Robusta coffee of the world, in Brazil, geographically located between the 39°38' and 41°50' West longitude meridians and the 17°52' and 21°19' South latitude parallels. The vulnerability was characterized by the spatial and temporal variation of rainfall and rainfall seasonal pattern (based on 30 years of historical data), elements of climatic water balance, elevation and area planted with Robusta coffee. The choice of mitigation and adaptation were based on widely validated criteria. Overall, the results show that the vulnerability of Robusta coffee is related to low index of rainfall, the rainfall seasonability and the water deficiency. In the studied region, there is approximately 42% of some type of water vulnerability during the year, with a severe to medium scale;this vulnerability is very pronounced in regions farther away from the coast of the Atlantic Ocean, since for a year approximately 92% of them are water deficient. In addition, the data show that this distance from the ocean implies a reduction of 75% in the phases of water surplus not only. The strategies of greater potential for adaptation and mitigation are related to the planting of improved genotypes, utilization of polycultures systems, increasing plant density, the implementation of irrigation systems and the management of spontaneous plants.