Biocontrol of green and blue molds in postharvest satsuma mandarin using Bacillus amyloliquefaciens JBC36

In order to control postharvest rot of satsuma mandarin (mandarin) fruits, a new strain of Bacillus amyloliquefaciens JBC36 (JBC36) was isolated from rhizosphere of a mandarin orchard and tested for its suppression of decay due to green and blue molds caused by Penicillium digitatum and Penicillium italicum, respectively, and its mode of action was investigated. In addition, carnauba wax-based and paraffin oil-based coating formulations were developed to increase the control efficacy of the antagonist. The strain JBC36 at 10⁸ CFU mL^?1 inhibited incidence of green and blue molds on wounded mandarin fruits with control efficacies of 88 and 80.2%, respectively. Mycelial growth and spore germination of P. digitatum and P. italicum were strongly inhibited in the presence of JBC36 or antagonistic metabolites. In order to determine antifungal activity, three kinds of antibiotics were isolated by reverse-phase high performance liquid chromatography (RP-HPLC) and identified as lipopeptide families, iturin A, fengycin and surfactin by RP-HPLC and thin-layer chromatography (TLC) analysis. Fengycin was further identified as C16 fengycin A by liquid chromatography/electrospray ionisation-mass spectrometry (LC/ESI-MS) and mass spectrometry/mass spectrometry (MS/MS) analysis. Volatile organic compounds from the antagonist also reduced the mycelial growth of P. digitatum and P. italicum. Carnauba wax-based and paraffin oil-based coating formulations containing 10⁸ CFU mL^?1 of JBC36 efficiently decreased the incidence of green mold with control efficacy of 91 and 80.9%, respectively. Overall, the antagonistic rhizobacterium JBC36 is a promising biocontrol agent for use in preventing postharvest spoilage of mandarin fruits by green and blue molds.     

Directional degradation of β-chitin by chitinase A1 revealed by a novel reducing end labelling technique

A novel procedure for labelling the molecular ends of β-chitin crystals has been established. By introducing a hydrazide derivative of biotin at the reducing end of a chitin chain, followed by a specific interaction between biotin and streptavidin coupled with a colloidal gold particle, the chain directionality of β-chitin microcrystals could be directly visualized by transmission electron microscopy. This method allowed to certify the parallelism of the chitin chains in the β-chitin microcrystals, and also to label the reducing tips of β-chitin microcrystals degraded by Bacillus circulans chitinase A1. With these substrates, the labelling occurred only at their tapered tip, which indicates that the digestion of these crystals proceeded from their reducing end. The generalization of this new labelling method to other polysaccharide crystals is discussed.     

Delay of early fruitlet abscission by branch girdling in citrus coincides with previous increases in carbohydrate and gibberellin concentrations

Current evidence in citrus indicates that gibberellins (GAs) are main determinants of early fruit set while subsequent growth of developing fruits is mostly dependent upon carbohydrate availability. In this work, branch girdling performed at anthesis in Satsuma mandarin (Citrus unshiu (Mak.) Marc.) cv. Okitsu transitorily reduced early abscission rates (12–32 days after anthesis, DAA) delaying initially the process of natural fruitlet drop. The effects of girdling on growth, gibberellin (GA) and carbohydrate concentrations in developing ovaries and fruitlets were assessed during this initial growth stage (0–69 DAA). In girdled branches, abscission rate reduction was preceded by elevated concentrations of carbohydrate and GA in developing ovaries and fruitlets. Girdling at anthesis stimulated higher hexose (21 DAA) and starch (6–20 DAA) concentrations and also higher GA₁ (6 DAA), GA₁₉ (13–20 DAA) and GA₂₀ (6–20 DAA). The results established a relationship between the reduction of early abscission rates and higher concentrations of carbohydrates and GAs induced by girdling in developing fruitlets. These findings revealed that girdling certainly increased GA concentration and strongly suggested that its effect on early fruitlet abscission delay is likely mediated by both GA and carbohydrates.     

Fatty acids of monogalactosyl diglycerides from Citrus

Fatty acids in vesicular and leaf monogalactosyl diglycerides (MGDG) of citrus were studied. Vesicular MGDG contained front 94.4 to 97.3% C₁₆, C_16:1, C_18:1, C_18:2, and C_18:3; whereas leaf MGDG contained ca 90% C_18:3, 3% C₁₆ and 1.8 to 9.5% C_18:2. Species varied considerably in their percentages of vesicular C_18:2, C1_8:3 and to a lesser degree, C_16:1 and C_18:1 fatty acids with lemons being the most distinctive. Branched fatty acids were present to the extent of 5.6% in vesicular and to only 0.1% in leaf MGDG.     

柑橘果实的光合特性、产物运输及分配在糖分积累中的作用

测定了温州蜜柑 (CitrusunshiuMarc .cv .Miyagawawase)果实发育进程中干鲜重、果皮光合速率和叶绿素含量的变化 ,并用14 CO2 示踪技术研究了果皮和叶同化生成的光合产物在果实内的运输分配特性。结果表明 :果皮光合速率与叶绿素含量有关 ,随着叶绿素含量的下降 ,果实光合速率也快速下降。在果实完熟之前 ,即使是当果皮积累的干重超过汁囊时 ,叶同化产物仍主要分配到汁囊中 ;而在完熟阶段 ,果皮光合速率接近零 ,果皮成了叶同化产物的主要库。果皮的同化产物 ,主要保留在果皮中 ,输入到汁囊的比率随果实发育而下降 ,但高峰时也有 12 %输入汁囊。与对照相比 ,果实遮光处理后降低了果皮与汁囊的干重和含糖量。上述结果表明果皮光合产物主要用于果皮自身的发育并能减少对叶光合产物的依赖 ,同时也能部分增加汁囊糖的积累     

柑橘果实的光合特性、产物运输及分配在糖分积累中的作用

测定了温州蜜柑(Citrus unshiu Marc. cv. Miyagawa wase)果实发育进程中干鲜重、果皮光合速率和叶绿素含量的变化,并用14CO2示踪技术研究了果皮和叶同化生成的光合产物在果实内的运输分配特性.结果表明:果皮光合速率与叶绿素含量有关,随着叶绿素含量的下降,果实光合速率也快速下降.在果实完熟之前,即使是当果皮积累的干重超过汁囊时,叶同化产物仍主要分配到汁囊中;而在完熟阶段,果皮光合速率接近零,果皮成了叶同化产物的主要库.果皮的同化产物,主要保留在果皮中,输入到汁囊的比率随果实发育而下降,但高峰时也有12%输入汁囊.与对照相比,果实遮光处理后降低了果皮与汁囊的干重和含糖量.上述结果表明果皮光合产物主要用于果皮自身的发育并能减少对叶光合产物的依赖,同时也能部分增加汁囊糖的积累.     

Production of somatic hybrids between satsuma mandarin (Citrus unshiu) and navel orange (Citrus sinensis) by protoplast fusion

We have regenerated altotetraploid plants that are interspecific somatic hybrids between Citrus sinensis Osbeck cv. Yoshida navel orange and Citrus unshiu Marc cv. Okitsu satsuma mandarin. Protoplasts isolated from ‘Yoshida’ leaves were chemically fused with call us-derived protoplasts from ‘Okitsu’. After 6 months of culture, 102 plants were obtained. These hybrids were identified by differential leaf morphology, DNA fluorescence intensity, and DNA analysis. Ploidy analysis via the flow cytometry revealed that 15 of the 102 plants were tetraploids, with the rest being diploids that morphologically resembled their mesophyll parent. SRAP analysis confirmed that 9 of the tetraploid plants were allotetraploid somatic hybrids. These will be utilized as a possible pollen parents for improving seedy citrus cultivars, e.g., ponkan, mandarin, lemon and kumquat, in order to produce triploid seedless hybrids.     

Antagonistic Changes between Abscisic Acid and Gibberellins in Citrus Fruits Subjected to a Series of Different Water Conditions

The relationship between absicisic acid (ABA) and gibberellin (GA) changes in developing fruitlets from both Clementina (Citrus clementina, Hort ex Tan) and Okitsu (Citrus unshiu, (Mak) Marc.) trees subjected to changing water conditions was investigated. The treatments consisted of a series of water stress, rainfall, and re-irrigation periods. To confirm the effectiveness of the imposed water changes, leaf water potential and soil moisture were measured. The data indicated that there were antagonistic changes between ABA and GA₂₀, because in both species ABA increased and GA₂₀ decreased during water stress, whereas re-hydration via either rainfall or irrigation reduced ABA but increased GA20. In addition, the data indicated that during water stress GA1 also decreased, whereas GA₈ did not change. After re-hydration, however, levels of GA₂₀ products, in general were rather dependent upon the hormonal levels induced in the previous water status. In conclusion, the results showed the occurrence of antagonistic changes between the levels of ABA and GA₂₀ in developing citrus fruitlets subjected to changing water conditions. The data might suggest that gibberellin 20-oxidase is regulated by water stress in citrus fruits.