Low‐cost agricultural waste accelerates tropical forest regeneration

Lower‐cost tropical forest restoration methods, particularly those framed as win–win business‐protected area partnerships, could dramatically increase the scale of tropical forest restoration activities, thereby providing a variety of societal and ecosystem benefits, including slowing both global biodiversity loss and climate change. Here we describe the long‐term regenerative effects of a direct application of agricultural waste on tropical dry forest. In 1998, as part of an innovative agricultural waste disposal service contract, an estimated 12,000 Mg of processed orange peels and pulp were applied to a 3 ha portion of a former cattle pasture with compacted, rocky, nutrient‐poor soils characteristic of prolonged fire‐based land management and overgrazing in Área de Conservación Guanacaste, northwestern Costa Rica. After 16 years, the experimental plot showed a threefold increase in woody plant species richness, a tripling of tree species evenness (Shannon Index), and a 176% increase in aboveground woody biomass over an adjacent control plot. Hemispheric photography showed significant increases in canopy closure in the area where orange waste was applied relative to control. Orange waste deposition significantly elevated levels of soil macronutrients and important micronutrients in samples taken 2 and 16 years after initial orange waste application. Our results point to promising opportunities for valuable synergisms between agricultural waste disposal and tropical forest restoration and carbon sequestration.     

Pathological categorization of the stem‐pitting and mild strains Citrus tristeza virus in Taiwan and their genomic analysis

Citrus tristeza virus (CTV), the causal agent of tristeza disease, causes the devastating diseases worldwide. In Taiwan, complex cultivars and long‐term infection by CTV result in more than 90% of infected citrus trees, but local strain identification and classification are still incomplete. Here, six CTV strains were categorized by grafting onto eight citrus cultivars and the pathological characteristics of the stem‐pitting mild strains were identified. After 6 months of inoculation, the pummelo stem‐pitting severe strain (CTV‐Pum/SP/T1) only caused severe symptoms in Wentan pummelo (WP) and the mild strain (CTV‐Pum/M/T5) was symptomless in every cultivar; the sweet orange (SO) stem‐pitting severe strain (CTV‐SwO/SP/T7) affected SO, WP and Ponkan mandarin (PM), and the mild strain (CTV‐SwO/M/T51) caused no symptoms in SO except for WP; the mandarin stem‐pitting severe strain (CTV‐Man/SP/T46) caused severe impacts in PM, WP and Eureka lemon, whereas the mild strain (CTV‐Man/M/T2) only caused severe stem‐pitting in WP. The full‐length sequencing of both pummelo stem‐pitting strains and phylogenetic analysis revealed that CTV‐Pum/SP/T1 and CTV‐Pum/M/T5 were related to the HA18‐9 and HA16‐5 strains from Hawaii, respectively. Moreover, recombination analysis revealed that TCT repeat sequences existed at open reading frame 1a in both the CTV‐Pum/SP/T1 and the T36 strains from the United States, indicating that the possible evolution relationship between two regions. Furthermore, improved universal and specific primer pairs were designed for more specific, sensitive detection to meet the needs for quarantine and early prevention. The understanding of strain pathogenicity and genomic analysis provided further characterization of each strain and enabled practical challenge inoculation against CTV disease.     

Comparative evaluation of petitgrain oils from six Citrus species alone and in combination as potential functional anti-radicals and antioxidant agents*

Six Citrus petitgrain oils, namely C. × sinensis (L.) Osbeck., C. × limon (L.) Osbeck., C. × clementina Hort., C. × aurantium L., C. reticulata Blanco and C. × bergamia Risso & Poit. (P1-P6), were characterized by means of gas chromatography equipped with flame ionisation detector (GC-FID) and gas chromatography coupled to mass spectrometry (GC-MS) and evaluated for their food functional-related properties. Antioxidant and radical-scavenging properties were tested by means of 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay, β-carotene bleaching test and Ferric Reducing Antioxidant Power (FRAP) assays. The synergistic interactions on antioxidant efficacy of petitgrain oils in combination were studied. The Relative Antioxidant Capacity Index (RACI) was applied from the perspective of statistics by integrating the antioxidant capacity data determined by all used methods. A high content of monoterpenes and a less representative content of sesquiterpenes characterized Citrus petitgrain oils. In the DPPH assay, C. × aurantium showed major effectiveness with an IC₅₀ value of 27.2 μg/ml. Citrus petitgrains oils, a rich source of healthy phytochemicals, should be considered a by-product of interest.     

Effect of lemon essential oil on the developmental stages of Trialeurodes vaporariorum West (Homoptera: Aleyrodidae)

Trialeurodes vaporariorum (Homoptera: Aleyrodidae), is a destructive pest of many ornamental and greenhouse crops throughout the world. In this research, effect of essential oil derived from lemon peel, Citrus aurantifolia (Hook.) on mortality of eggs, first-instar nymphs and on adult oviposition of T. vaporariorum (Westwood) was determined under laboratory conditions. Analysis of Citrus aurantifolia essential oil used for insect fumigation by phase gas chromatography revealed the presence of 16 compounds including Limonene (56.6%), β-pinene (16.3%) and α-terpineol (11.3%). Five concentrations of essential oil – 0.002, 0.004, 0.008, 0.016 and 0.032 (μl/ml) – were applied in fumigant toxicity experiments. Greater mortality was observed with increasing dose of essential oil. First-instar nymphs were more sensitive to essential oil treatments compared with eggs and adults that reduced the survival rate of T. vaporariorum by 58, 70 and 56% after treatment of eggs, nymphs and adult, respectively. Based on this study, essential oil derived from C. aurantifolia could be used as an effective and environmentally sustainable bioinsecticide for the control of T. vaporariorum.     

Purification of an Iron-Containing Superoxide Dismutase from A Citrus Plant,Citrus Limonum R

A cyanide-insensitive superoxide dismutase was purified to apparent homogeneity from lemon leaves (Citrus limonum R). The enzyme was isolated from leaf extracts by ammonium sulfate salting-out. and ion-exchange, gel filtration and hydroxylapatite column chromatography. The purified Fe-SOD had a specific activity of about 1.500 U/mg and represents approximately 1.6% of the total soluble protein in lemon leaf extracts. A molecular weight of 47,500 was determined for the enzyme. Analytical gel electro-focusing of the purified preparation revealed the presence of two isozymes with pl values of 5.13 and 4.98. Metal analysis showed the presence of I g-atom of iron and 0.5g-atom of manganese per mol of enzyme. The visible and UV absorption spectra of the Citrus enzyme were similar to those reported for other iron-containing SODS from different origins. The significance of the presence of Fe-SOD in higher plants is briefly discussed.     

Embriogenesi nucellare in ovuli non sviluppati di alcune cultivar di arancio del gruppo navel

Abstract

Nucellar embryogenesis in undeveloped ovules in some navel orange cultivars. - Nucellar embryogenesis has been studied in some cultivars of navel orange, Citrus sinensis (L.) Osbeck. Histological observations have been carried out in undeveloped ovules excised 2, 4, 6, 8 months after anthesis. Ovules under study derived a) from flowers naturally pollinated or hand-pollinated with pollen of Poncirus trifoliata (L.) Raf.; b) from emasculated flowers, and c) from flowers deprived of stigma. In absence of pollination, as well as in absence of fertilization, the nucellar embryos reached the globular stage.     

Isotope Ratio by HRGC-MS of Citrus junos Tanaka (Yuzu) Essential Oils: m/z 137/136 of Terpene Hydrocarbons

The isotope ratios of monoterpene hydrocarbons in Citrus junos Tanaka (yuzu) essential oils from different origins were determined by ordinary high-resolution gas chromatography-mass spectrometry (HRGC-MS). Both intensities of the molecular mass peaks (m/z 136) and of the isotope peaks (m/z 137) of monoterpene hydrocarbons were measured by single-ion monitoring with an MS analysis. The isotope ratios (m/z 137/136) of the ten monoterpene hydrocarbons commonly contained in citrus essential oils, α-pinene, β-pinene, sabinene, myrcene, α-phellandrene, α-terpinene, limonene, γ-terpinene, β-phellandrene and terpinolene, were determined in yuzu samples of the highest commercial quality from 42 different production districts. Statistical treatment of these data by the t-test and sign test revealed significant differences of the isotope effects in each yuzu sample. It is suggested that this technique will be applicable for evaluating the quality, genuineness and origin of citrus fruits and their products. The isotope fingerprints were also demonstrated in several citrus fruits other than the yuzu samples.     

Xanthan is not essential for pathogenicity in citrus canker but contributes to<Emphasis Type="Italic"> Xanthomonas</Emphasis> epiphytic survival

Xanthan-deficient mutants of Xanthomonas axonopodis pv. citri, the bacterium responsible for citrus canker, were generated by deletion and marker exchange of the region encoding the carboxy-terminal end of the first glycosyltransferase, GumD. Mutants of gumD did not produce xanthan and remained pathogenic in citrus plants to the same extent as wild-type bacteria. The kinetics of appearance of initial symptoms, areas of plant material affected, and growth of bacteria inside plant tissue throughout the disease process were similar for both wild-type and mutant inoculations. Moreover, exopolysaccharide deficiency did not impair the ability of the bacteria to induce hypersensitive response on non-host plants. Apart from variations in phenotypic aspects, no differences in growth or survival under different stress conditions were observed between the xanthan-deficient mutant and wild-type bacteria. However, gumD mutants displayed impaired survival under oxidative stress during stationary phase as well as impaired epiphytic survival on citrus leaves. Our results suggest that xanthan does not play an essential role in citrus canker at the initial stages of infection or in the incompatible interactions between X. axonopodis pv. citri and non-host plants, but facilitates the maintenance of bacteria on the host plant, possibly improving the efficiency of colonization of distant tissue.     

化橘红果实生长发育过程中类黄酮的动态变化

该研究建立了化橘红(Citrus grandis cv.‘Tomentosa’)果实中4种类黄酮成分的提取分离、HPLC定量和指纹图谱分析方法,利用该方法测定了果实生长发育过程中类黄酮的动态变化。结果表明,用60%乙醇超声辅助提取果实干燥粉末2小时,柚皮苷提取率达到98%以上。用HPLC分离和测定提取物中柚皮苷、野漆树苷、柚皮素和芹菜素的含量及其指纹图谱。结果显示,15–60天不同果龄的果实中类黄酮含量(占果实干重)随着果龄的增大而降低,柚皮苷、野漆树苷、柚皮素和芹菜素分别从52.5%、0.74%、0.57%和0.23%降低至16.1%、0.11%、0.06%和0.03%;每果中类黄酮总量则随着果龄的增加而大幅提高,从15天的0.55 g提高至60天的7.99 g。上述研究结果表明,果龄对化橘红类黄酮含量、产量及药材质量均有很大的影响。该研究为化橘红的工业生产和质量监控提供了重要依据。