Sinensis Trees Research Articles

Grafted twig rot on Citrus sinensis caused by a member of the Fusarium solani species complex

Citrus sinensis is an important fruit crop valued for its juicy fruit pulp and aromatic rind. Grafted twigs with rot progressing from both ends were observed on C. sinensis trees in 12 orchards in Zigui, China, in June 2017, with only 5% of grafted twigs surviving in some orchards. Severe infection resulted in a rot of the entire twig, with the twig becoming covered by a white mycelium. A fungus frequently recovered from the diseased twigs was identified as Fusarium solani based on cultural and conidial characteristics. Multi-gene phylogenetic analysis including partial gene sequences of the internal transcribed spacer (ITS) region of rDNA, translation-elongation factor 1 alpha (EF-1ɑ) and the RNA polymerase second largest subunit (RPB2) indicated that the isolated fungus belonged to the Fusarium solani species complex (FSSC) and was related closely to FSSC22 recovered from Xanthoxylum piperitum in Japan. Pathogenicity tests confirmed that this fungus is the causal agent of grafted twig rot on C. sinensis.

Mass spectrometry imaging guided molecular networking to expedite discovery and structural analysis of agarwood natural products

Structural analysis of biomolecules is essential to natural product discovery, especially for precious biomaterials such as agarwood. However, one of the greatest challenges to the characterization of natural products is the profound cost in time and manpower to the structural elucidation of these highly diverse compounds. Here, we demonstrate a multi-modal mass spectrometric strategy, integrating matrix-assisted laser desorption ionization (MALDI) mass spectrometry imaging (MSI) and mass spectral molecular networking, to uncover agarwood natural products of Aquilaria sinensis trees. A simple workflow for preparing wood sections for MALDI-MSI analysis was demonstrated. Notably, tens of natural products in the agarwood region in wood stem section of A. sinensis were spatially revealed by MALDI-MSI. For the first time, such a great number of plant specialized metabolites is obtained by a single wood section MSI. Guided by the spatially resolved features, mass spectral molecular networking was subsequently applied for structural analysis of the agarwood natural products, in which three major classes of 2-(2-phenylethyl)chromones and their analogues were putatively characterized. These results suggest an efficient strategy to the dereplication of plant natural products.

Dynamic analysis of gene expression and determination of chemicals in agarwood in Aquilaria sinensis

Agarwood is the resinous heartwood of Aquilaria species. However, low yields and high costs of existing stimulation methods have led to the need for new techniques to produce agarwood rapidly and effectively. We developed a biological agarwood-inducing technique (Agar-Bit) that produces high yields and quality within 6 months. To better understand agarwood formation by Agar-Bit, dynamic gene expressions of key synthetases pathways of sesquiterpenes and chalcone- related enzymes at different times were determined after both Agar-Bit and the traditional burning chisel drilling (BCD) stimulation on Aquilaria sinensis trees. The qRT-PCR results show that some characteristic synthase genes were expressed at greatly different levels and times compared with the controls. For the Agar-Bit technology, main changes were after the 3rd or 5th month, while BCD expression clearly changed at the 5th month. Essential oils and total chromone contents were simultaneously determined. In the Agar-Bit group, both were higher and similar to natural levels. The Agar-Bit methodology is a new option for producing agarwood as demonstrated by genetic and chemical aspects. The differences in gene expression within 6 months for both groups indicates that the mechanisms of the two methods are different. These findings provide information on genetic variation during the process of agarwood formation.

GC-MS Study of the Chemical Components of Different Aquilaria sinensis (Lour.) Gilgorgans and Agarwood from Different Asian Countries.

As a traditional medicinal herb and valuable natural spice in China, Aquilaria sinensis (Lour.) Gilg has many significant pharmacological effects. Agarwood is the resinous heartwood acquired from wounded A. sinensis trees, and is widely used in pharmaceuticals owing to its excellent medicinal value. In this study, the chemical composition of volatile components and alcohol extracts from different organs of A. sinensis and agarwoods grown in different regions were investigated using GC-MS. The results showed that Vietnam agarwood had the highest moisture content, which was attributed to the local climate, while the fruit and bark of A. sinensis had higher moisture contents than the other organs. The volatile components of A. sinensis organs included 3-ethyl-5-(2-ethylbutyl)-octadecane, oleic acid 3-(octadecyloxy) propyl ester, and docosanoic acid 1,2,3-propanetriyl ester, while the alcohol extracts of A. sinensis organs contained benzoic acid ethyl ester, hexadecanoic acid ethyl ester, oleic acid, and n-hexadecanoic acid. Furthermore, the main active ingredients in agarwood from different habitats were sesquiterpenoids, aromatic species, and chromone compounds. The role of chromone compound 2-phenylethyl-benzopyran as an elicitor and the mechanism of agarwood formation were also investigated. Antioxidant tests showed that essential oils from agarwood and A. sinensis had antioxidant capacities by comparison with butylated hydroxytoluene and vitamin E. An antibacterial activity test showed that the inhibition effect of the essential oil was better against Gram-positive bacteria than against Gram-negative bacteria.

Trunk surface agarwood-inducing technique with Rigidoporus vinctus: An efficient novel method for agarwood production.

Only when Aquilaria spp. or Gyrinops spp. trees are wounded, due to insect attack, or microbial invasion, agarwood can be successfully induced. In the present study, a fungus which can induce agarwood formation efficiently was isolated and a suitable method for its application to induce agarwood formation was developed. Rigidoporus vinctus was isolated from the inner layers from infectious A. sinensis trees. When the fermentation liquid of fungi inoculated back to A. sinensis tree, agarwood was found to be induced. In addition, a novel method called trunk surface agarwood-inducing technique (Agar-Sit) was developed to produce agarwood with R. vinctus. The alcohol soluble extract content of the agarwood, up to 38.9%, far higher than the requirement (10%) in Chinese Pharmacopoeia and the six characteristic compounds of agarwood used as Chinese Medicinal Materials were all detected. Their relative percentages of the sesquiterpenes in the essential oil were 22.76%. This is the first report of the Agar-Sit and also the application of R. vinctus in agarwood induction. According to the results, when the combination of Agar-Sit and R. vinctus is used agarwood can be induced with high yield and good quality.

Harvesting Method Affects Water Dynamics and Yield of Sweet Orange with Huanglongbing

Changes in grove management practices may change crop water dynamics. The objective of this study was to estimate sap flow, stem water potential (Ψstem), and citrus yield as affected by harvesting methods in sweet orange (Citrus sinensis) trees affected by Huanglongbing. The study was initiated in March 2015 for two years on five-year-old commercial sweet orange trees at a commercial grove located at Felda, Florida (26.61° N, 81.48° W) on Felda fine sand soil (Loamy, siliceous, superactive, hyperthermic Arenic Endoaqualfs). All measurements were replicated before and after harvest in four experiments (A, B, C and D) under hand and mechanical harvesting treatments. Sap flow measurements were taken on four trees per treatment with two sensors per tree. Sap flow measured by the heat balance method at hourly intervals during March and April of 2015 and 2016 significantly declined after harvesting by 25% and 35% after hand and mechanical harvesting, respectively. Ψstem measured after harvest was significantly higher than measurements before harvest. The average value of Ψstem measured increased by 10% and 6% after hand and mechanical harvesting, respectively. Mechanical harvesting exhibited lower fruit yields that averaged between 83%, 63%, 49% and 36% of hand-harvested trees under A, B, C and D experiments, respectively. It is concluded that the hand harvesting method is less stressful and less impactful on tree water uptake and fruit yield compared with mechanical harvesting.

Severity and genetic diversity of Citrus tristeza virus complexes infecting ‘Pêra Bianchi’ sweet orange in Paraná, Brazil

The aggressiveness and genetic diversity of the Citrus tristeza virus (CTV) infecting sweet orange ‘Pera Bianchi’ (Citrus sinensis) trees were evaluated under the environmental conditions of the state of Parana, Brazil. Disease severity was assessed based on stem pitting symptoms in the citrus trees, while genetic diversity was determined by sequencing the virus coat protein (CP) gene. Stem pitting symptoms were less severe in trees maintained under greenhouse conditions and field experimental plots in the municipalities of Paranavai and Umuarama, where the temperature is higher. In contrast, the most severe stem pitting symptoms were observed in trees from experimental orchards established in colder areas (i.e. municipalities of Arapongas, Cascavel and Londrina), suggesting a temperature effect on symptom development. Analysis of CP sequences indicated intra- and interspecific diversity of the CTV complex for most of the trees included in this study. Recombination analyses of the CP gene also allowed to identify recombinants with parentals related to PIAC-5, a mild isolate, and 1BL-3, found in trees from a field plot in Londrina. The CTV complex infecting ‘Pera Bianchi’ trees under field conditions showed a high degree of genetic diversity, implying the possibility of multiple infections or even a dynamic rearrangement of the genotypes into different CTV complexes under the environmental conditions of the state of Parana.

Agarwood Formation Induced by Fermentation Liquid of Lasiodiplodia theobromae, the Dominating Fungus in Wounded Wood of Aquilaria sinensis

Agarwood is broadly used in incense and medicine. Traditionally, agarwood formation is induced by wounding the trunks and branches of some species of Aquilaria spp., including A. sinensis. As recently evidenced, some fungi or their fermentation liquid may have the potential of inducing agarwood formation. The present study aimed to analyze the fungi isolated from an agarwood-producing A. sinensis tree and subsequently identify the fungi capable of promoting agarwood formation. We identified a total of 110 fungi isolates based on their morphological characteristics and rDNA ITS sequences. These isolates came from four different layers (namely the decomposing layer, agarwood layer, transition layer, and normal layer) near the agarwood formation site of the trunk. According to the experimental results, most of them belonged to Dothideomycetes (81.82%), while the others to Sordariomycetes (13.64%) or Eurotiomycetes (4.55%). Of note, 88 isolates were shown belonging to the species of Lasiodiplodia theobromae that are most frequently isolated from different layers. In addition, when the fermentation liquid of two isolates of L. theobromae (AF4 and AF12) and one isolate of Fusarium solani (AF21) was inoculated into the A. sinensis wood using the Agar-Wit technique, promoted agarwood formation was observed; however, the effect of AF21 did not keep stable in the later test, while AF4 and AF12 still functioned 1 year later. This study may lay a foundation for exploring the underlying mechanism of agarwood formation as well as fungi application in agarwood production.

Diversity of endophytic bacteria from Citrus and Inga trees in the coffee agro-ecosystem and fluctuations by location, shade management and season

Coffee, Coffea arabica L., is planted in diverse agro-ecological environments in the mountains of Puerto Rico (PR). The pathogen, Xylella fastidiosa Wells et al., is a xylem-limited phytopathogenic bacterium transmitted by insects that causes coffee leaf scorch, citrus-variegated chlorosis and vascular diseases of many other plant species. The objective of this study was to characterize the fastidious endophytes occupying a similar niche as X. fastidiosa in the vascular system of Citrus sinensis and Inga vera trees in low diversity and high diversity shade environments where coffee is grown In PR. Sampling was conducted over two years in four localities: Adjuntas, Jayuya, Las Marías and Yauco during the dry and rainy seasons. Citrus sinensis and Inga vera were generally the only shade treesin coffee plantations described as "low diversity"; in "high diversity" plantations these same species were present along with Pithocellobium carbonarium, Gliricidia sepium and Andira inermis. Endophyte population (EP) and endophyte diversity (ED) were contrasted using factorial arrangements of shade, locality, season and year. Endophytes were clustered using multivariate analysis. Neither shade nor season had an effect on EP in Citrus. In Inga vera, EP was about three times higher in the second year compared to the first year regardless of season, location or shade. In both Citrus and Inga, ED was low. The highest ED was detected in Jayuya and the lowest in Las Marías. Ward's algorithm combined Citrus and Inga fastidious strains into four and five clusters for Gram-negative and Gram-positive bacteria, respectively. Endophyte diversity of Citrus and Inga was determined by the plant species, not by location or shade. However, ED was higher in shaded coffee with low diversity of tree species. Thus, effects of shade (low versus high diversity) altered the community structure of endophytes prevalent in the vascular tissue and may impact host plant resistance to pathogens such as X. fastidiosa.

Transcriptome Sequencing of Chemically Induced Aquilaria sinensis to Identify Genes Related to Agarwood Formation.

BackgroundAgarwood is a traditional Chinese medicine used as a clinical sedative, carminative, and antiemetic drug. Agarwood is formed in Aquilaria sinensis when A. sinensis trees are threatened by external physical, chemical injury or endophytic fungal irritation. However, the mechanism of agarwood formation via chemical induction remains unclear. In this study, we characterized the transcriptome of different parts of a chemically induced A. sinensis trunk sample with agarwood. The Illumina sequencing platform was used to identify the genes involved in agarwood formation.Methodology/Principal FindingsA five-year-old Aquilaria sinensis treated by formic acid was selected. The white wood part (B1 sample), the transition part between agarwood and white wood (W2 sample), the agarwood part (J3 sample), and the rotten wood part (F5 sample) were collected for transcriptome sequencing. Accordingly, 54,685,634 clean reads, which were assembled into 83,467 unigenes, were obtained with a Q20 value of 97.5%. A total of 50,565 unigenes were annotated using the Nr, Nt, SWISS-PROT, KEGG, COG, and GO databases. In particular, 171,331,352 unigenes were annotated by various pathways, including the sesquiterpenoid (ko00909) and plant–pathogen interaction (ko03040) pathways. These pathways were related to sesquiterpenoid biosynthesis and defensive responses to chemical stimulation.Conclusions/SignificanceThe transcriptome data of the different parts of the chemically induced A. sinensis trunk provide a rich source of materials for discovering and identifying the genes involved in sesquiterpenoid production and in defensive responses to chemical stimulation. This study is the first to use de novo sequencing and transcriptome assembly for different parts of chemically induced A. sinensis. Results demonstrate that the sesquiterpenoid biosynthesis pathway and WRKY transcription factor play important roles in agarwood formation via chemical induction. The comparative analysis of the transcriptome data of agarwood and A. sinensis lays the foundation for elucidating the mechanism of agarwood formation via chemical induction, and thus, enables future improvements in agarwood quality while protecting endangered wild A. sinensis.

First report of olive leaf scorch in Brazil, associated with Xylella fastidiosa subsp. pauca

Olea europaea (L.) trees displaying leaf scorching symptoms, identical to those recently reported for olive trees colonized by Xylella fastidiosa in Southern Italy and also in Argentina, were observed in commercial orchards of two counties in Southeastern Brazil. PCR-based diagnosis using conserved primers for X. fastidiosa strains (RST31/33) and also specific to X. fastidiaosa subsp. pauca (CVC1/272-2 int) were positive for all symptomatic tested samples (n = 8 of 9), but no template was obtained using twigs from asymptomatic trees (n = 20). Bacterial colonies were isolated from symptomatic tissues on culture medium and confirmed by PCR using the set of primers specific to X. fastidiosa subsp. pauca . Comparative sequence analyses of seven MLST loci amplified from one tripled passaged colony (MFG01) perfectly matched with sequences of alleles leuA #7, petC #6, malF #8, cysG #10, holC #11, nuoL #8, and gltT #8, the allelic profile of Sequence Type-ST16, which is represented by the strain COF0238 isolated from Coffea arabica (L.) in Brazil (http://pubmlst.org/xfastidiosa/). Phylogenetic analysis placed the ST16 into subspecies pauca, but genetically closer to ST11 and ST13, both obtained from Citrus sinensis (L.) trees with citrus variegated chlorosis. The results confirm the association of olive plants showing leaf scorching with the presence of X. fastidiosa subsp. pauca , and represent the first report of this bacterium in Brazilian olive orchards.

Effects of agarwood formation induced by Fusarium sp. A2 on distribution and community composition of endophytic fungi in leaves of Aquilaria sinensis

The aim of the paper is to observe the distribution of the endophytic fungi in leaves of Aquilaria sinensis by using permanent paraffin-cut section, optical microscope photography and histochemistry. Total DNA was extracted with modified CTAB method and rDNA ITS regions of plant and endophytic fungi were amplified with eukaryotic universal primers. The rDNA ITS amplicon was characterized by RFLP analysis, sequencing of rDNA ITS library and phylogenetic analyses using PAUP by maximum parsimony. Fusarium sp. A2 was used to induce the formation of resinous in A. sinensis trees. As a result, endophytic fungi mainly distributed in spongy and phloem in leaf. Endophytic fungi distributed in the phloem in agarwood-producing tree and had a relatively high abundance. Phoma sp. and Collectrotrichum sp. were the absolute advantage species in the leaf tissues of non-resinous and agarwood-producing tree, respectively. Collectrotrichum sp. was the only fungal species detected both in the two types of A. sinensis with different levels of abundance. The culture-independent molecular method can be used to identify fungal species directly and rapidly from the plant tissues. Endophytic fungal communities in non-resinous and agarwood-producing A. sinensis leaf tissues were quite different.

First Report of Branch Dieback of Walnut Caused by Neofusicoccum parvum in Korea.

Walnut (Juglans sinensis Dode) is an economically important tree in the world, both for its wood and its fruit. Walnut fruits, as rich sources of omega-3 essential fatty acid, are valuable nutritionally. Consumer interest in Korea for walnuts has increased in recent years, and production has increased to 1,042 ha with the Kyoungbuk region consisting of 402 ha (2). In May 2012, lethal dieback disease of walnut tree was detected in two orchards in Andong, Kyoungbuk region, Korea, each with an incidence of 25 to 30%. Disease symptoms included blight and dieback of the stems, flowing resin, dark decay inside the bark of dead twigs, and defoliation. The bark of dead twigs was removed and sliced thinly using a razor blade, and water-mounted, without staining, for observation of fungal structures, if present. Pycnidia were found embedded within the bark of dead twigs and conidia were mostly characterized by fusoid, hyaline, smooth, thin-walled, unicellular and 16.25 to 21.25 μm long and 4.37 to 6.87 μm wide. These characteristics are consistent with those reported previously for Neofusicoccum parvum (Pennycook & Samuels) Crous, Slippers & A.J.L. Phillips (1). Diseased branch tissues collected from the two locations were surface sterilized with 1% NaOCl, rinsed with sterile distilled water, and plated on potato dextrose agar (PDA). The fungal isolates, recovered from the two different orchards, produced white, aerial mycelium and became light gray within a week after incubating plates at 25°C. To confirm the identities of the isolates, the complete internal transcribed spacer (ITS) rDNA of the fungi was amplified and sequenced using PCR. The sequences were compared with other DNA sequences in the GenBank database, using a BLAST search. BLAST analysis of the PCR product showed that the sequence had 99% identity with the nucleotide sequences for N. parvum (JQ411396.1 and GU997688.1). Additionally, the chitin synthase 1 gene was sequenced and analyzed using the BLAST server. The sequence of PCR product had 100% identity with the nucleotide sequences of N. parvum strain CMW9080 chitin synthase 1 gene (EU339501). Thus, both morphological and molecular characters confirmed this species as N. parvum. Pathogenecity tests were performed by inoculating 2-year-old J. sinensis trees. Inoculations consisted of inserting 5-mm-diamter agar plug bearing fresh mycelium of the fungal isolates into the wounds. Within 2 weeks, black lesions appeared on all inoculated plants accompanied by defoliation, whereas no symptoms were observed in the control plants. N. parvum has been reported a member of Botryosphaeriaceae, commonly associated with dieback and cankers of woody plants (1). To our knowledge, this study is the first report of N. parvum as a pathogen of Juglans sinensis in Korea.

Effects of inoculating fungi on agilawood formation in Aquilaria sinensis

Agilawood is a costly heartwood medicine obtained from Aquilaria sinensis with active ingredients mainly composed of volatile and semi-volatile substances. However, the formation time of agilawood is quite long and little is known about its formation mechanism. Two highly active fungi obtained from natural agilawood were inoculated on A. sinensis trees to understand their interaction processes and elucidate the transformation rules of induced chemical compositions within different test periods. The results demonstrated that the fungi could successfully colonize living tissues and cells and activate the host defense system, resulting in agilawood accumulation. With increasing time, the main components of A. sinensis converted into constituents or analogs of agilawood and the host exhibited “self-injury” to prevent fungal intrusion and protect other tissues. The data presented here could provide scientific basis for producing agilawood with the two new fungi in a safe, feasible, and sustainable manner without destroying rare Aquilaria plants.

Molecular identification of endophytic fungi from Aquilaria sinensis and artificial agarwood induced by pinholes-infusion technique

Agarwood, the resinous portions of Aquilaria plants, have been used as medicines and incenses. Aquilaria sinensis is the major producer of agarwood in China. Agarwood are generally viewed as pathological products formed as defense symptom against fungal infection. In this study, microbial communities inhabiting the leaves of non-resinous and agarwood-producing wounded A. sinensis tree were investigated by cultivation-independent approaches, such as PCR, restriction fragment length polymorphism (RFLP) analysis and sequencing of rDNA internal transcribed spacer (ITS) library. Molecular phylogenetic analysis demonstrated that Botryosphaeria, Colletotrichum gloeosporioides, Phomopsis and Cylindrocladium species are members of the agarwood-producing wounded tree, while Phoma, Mycosphaerella, Sagenomella, Alternaria and Ramichloridium species is able to colonize the non-resinous tree internally. C. gloeosporioides was the only fungus shared by the two rDNA ITS libraries. C. gloeosporoides, Botryosphaeria , and Cylindrocladium were considered to be related to agarwood formation. A pinholes-infusion method to induce the generation of agarwood by chemically stimulated and/or inoculate combined method was established. One to two years after the artificial inoculation, resinous wood were collected and the inoculating effects were detected by ethanol extraction content, thin layer chromatography (TLC) and gas chromatography-mass spectrometry (GCMS) techniques. The results reveal that chemically stimulated with formic acid and infected by Botryosphaeria dothidea produced high yield and high quality artificial agarwood in a relatively short time. Keywords : Agarwood, endophytic fungi, Aquilaria sinensis , molecular identification, artificial induce of agarwood African Journal of Biotechnology Vol. 12(21), pp. 3115-3131

Whole-tree Agarwood-Inducing Technique: An Efficient Novel Technique for Producing High-Quality Agarwood in Cultivated Aquilaria sinensis Trees

Agarwood is the fragrant resin-infused wood derived from the wounded trees of Aquilaria species. It is a valuable non-timber forest product used in fragrances and as medicine. Reforestation for Aquilaria trees in combination with artificial agarwood-inducing methods serves as a way to supply agarwood and conserve of wild Aquilaria stock. However, the existing agarwood-inducing methods produce poor-quality agarwood at low yield. Our study evaluated a novel technique for producing agarwood in cultivated Aquilaria trees, called the whole-tree agarwood-inducing technique (Agar-Wit). Ten different agarwood inducers were used for comparison of Agar-Wit with three existing agarwood-inducing methods. For Aquilaria trees treated with these ten inducers, agarwood formed and spread throughout the entire tree from the transfusion point in the trunk to the roots and branches of the whole tree. Agarwood yield per tree reached 2,444.83 to 5,860.74 g, which is 4 to 28 times higher than that by the existing agarwood-inducing methods. Furthermore, this agarwood derived from Agar-Wit induction was found to have a higher quality compared with the existing methods, and similar to that of wild agarwood. This indicates Agar-Wit may have commercial potential. Induction of cultivated agarwood using this method could satisfy the significant demand for agarwood, while conserving and protecting the remaining wild Aquilaria trees.

Avaliação genética de seleções e híbridos de limões cravo, volkameriano e rugoso como porta-enxertos para laranjeiras Valência na presença da morte súbita dos citros

Este estudo teve como objetivo realizar a avaliação genética da produção de frutos, eficiência produtiva e altura de laranjeiras Valência (Citrus sinensis) enxertadas em seleções e híbridos dos limões Cravo (C. limonia), Volkameriano (C. volkameriana) e Rugoso (C. jambhiri), em área endêmica para morte súbita dos citros (MSC). Foram avaliados 36 genótipos desses porta-enxertos, representados por cinco plantas cada, avaliados em cinco safras, do terceiro ao sétimo ano após o plantio. Sete dos genótipos avaliados apresentaram plantas com sintomas de MSC até o sétimo ano: Rangpur Otaheite orange 12901 (859), Rangpur Red Lime D.33.30 (866), Limão-Cravo EEL (871), Rangpur Borneo red (874), Citrus kokhai (1649), Limão-Rugoso 58329 (1655) e Limão- Cravo x Swingle B (1695). Para os genótipos que não manifestaram sintomas da doença, foram estimados parâmetros genéticos e fenotípicos e realizada a predição de valores genéticos dos indivíduos, visando à seleção e ao melhoramento genético para as características citadas, empregando-se o método REML/BLUP (máxima verossimilhança restrita/melhor predição linear não viciada). A análise de produção de frutos de cinco safras mostrou acurácia seletiva de 84,59%, tornando-se desnecessária a avaliação de maior número de safras. A seleção dos sete melhores genótipos proporcionou ganhos genéticos de 11,5% na produção de frutos, enquanto a do melhor genótipo conferiu ganho genético de 16,3%. As maiores médias genéticas preditas (>70,0 kg.pl-1) para produção de frutos foram obtidas pelos genótipos Limão-Cravo- Ipanema (1522), Santa- Bárbara-Red- Lime (884), Limão- Cravo- Limeira (863), Limão- Cravo- Taquaritinga (869), Limão- Rugoso- do -Cabo (1643), Rangpur- Rose Lime (868) e Limão- Cravo- da- Califórnia (1467). Já a acurácia seletiva da eficiência produtiva, para quatro colheitas, foi 77,4%. Para este caráter, as maiores médias genéticas (>8,0 kg.m-3) foram dos genótipos Rangpur- Lime x Trifoliata 3810 (1648), Rangpur- Lime x Trifoliata 5320 (1644), Limão- Cravo x Citrange- Carrizo (1524), Citrus pennivesiculata (880), Limão- Cravo x Trifoliata- Swingle A (1707), Rangpur- Rose- Lemon 124684 (864), Rangpur- Red -Lime D33.47 (867) e Limão- Cravo -Ipanema (1522). Dentre os 10 melhores genótipos para produção de frutos e para eficiência produtiva, apenas três são coincidentes: Rangpur- Rose -Lime (868), Citrus pennivesiculata (880) e Limão- Cravo-Ipanema (1522).

Historical records and modern studies on agarwood production method and overall agarwood production method

Agarwood is a precious traditional Chinese medicine with the efficacy of promoting qi circulation and relieving pain, warming middle-jiao, controlling nausea and vomiting, governing inspiration and relieving asthma, therefore it is widely applied in the clinic. Meanwhile, agarwood is also a precious spice. Aquilaria sinensis is the only source of agarwood production in China. Under natural conditions, a healthy A. sinensis tree produces no agarwood. Only if being wounded or infected with fungus can it synthetize and accumulate agarwood. It takes a decade or even several decades to produce agarwood, thus natural agarwood can not meet market demands. The essay summarizes historical records of agarwood production method and modern agarwood production method, in order to provide basis and reference for large-scale production of agarwood.

Production of high-quality agarwood in Aquilaria sinensis trees via whole-tree agarwood-induction technology

We used whole-tree agarwood-induction technology to produce agarwood from Aquilaria sinensis trees within 20 months, and evaluated the quality of this agarwood. The results showed its characteristics were similar to those of high-grade wild agarwood in terms of texture, chemical constituents, essential oil content, and ethanol-soluble extract content, with the lattermost quality far surpassing the requirement of traditional Chinese medicine agarwood, as indicated in Chinese Pharmacopoeia 2010. To the best of our knowledge, this is first study to show that high-quality agarwood can be produced in whole A. sinensis trees via a chemically induced technology.

Determination of Bioactive Components of Waste Leaves from Chinese Hackberry by GC/MS

Chinese Hackberry (Celtis sinensis) tree has high adaptability to environment and tolerance to water stress, and a long history of utilization and plantation in many countries. However, researches on Chinese Hackberry were mostly focused on the biomass analyses and utilizations of its wood, and lacked those to analyze the chemical components of extractives of Chinese Hackberry leaves, which was very important to recover and utilize the polluting waste Chinese Hackberry leaves. Therefore, the chemical components of benzene/ethanol extractives of Chinese Hackberry leaves were analyzed by method of GC/MS in order to identify top value-added bioactive components from waste leaves of Chinese Hackberry tree. The analytical result showed that the main components of benzene/ethanol extractives of freeze-dried Chinese Hackberry leaves by GC/MS analysis were identified 10 components (106 peaks) as: Sesquirosefuran (48.57%), Thiophene, 2-ethyltetrahydro- (20.01 %), Hexatriacontane (8.67%), Octadecane (8.60%), 1-Eicosanol Pregn-4-en-3-one, 20-hydroxy-, (20R)- (4.52%), 1-Amino-2-(hydroxymethyl)anthraquinone (3.63%), 1,3-Diphenyl-(4H)1,2,4-triazoline- (1.92%), Tricyclo[4.3.1.13,8]undecane-1-carboxylic acid (1.69%), Eucalyptol (1.60%), etc. As the first report here, our result by GC/MS showed that the benzene-methanol extractive of freeze-dried Chinese Hackberry leaves can be developed into top value-added materials of spicery, biomedicines, and biofuel.