Physical mapping of the 18S-25S and 5S ribosomal RNA genes in diploid bananas

Fluorescent in situ hybridisation (FISH) was used to determine the number and distribution of the 18S-25S and 5S rDNA sites on mitotic chromosomes of 6 wild and 2 edible diploid (2n=22) accessions belonging to the two banana species, Musa acuminata and M. balbisiana. FISH with the 18S-25S probe resulted in signals on one pair of chromosomes, the position of signals corresponded to the secondary constriction at the end of a short arm. The intensity of labelling was different between the homologues and the larger site corresponded to a larger secondary constriction. This labelling pattern was observed consistently in all genotypes. On the other hand, differences in the number of 5S sites were observed between the accessions. While in some of the wild seeded species, the 5S rDNA was localised on two pairs of chromosomes, hybridisation signals appeared on three pairs of chromosomes in other wild accessions. Quite unexpectedly, only five sites of 5S rDNA were reproducibly observed in the two vegetatively propagated diploid edible cultivars, Pisang Mas and Niyarma Yik, evidence for structural heterozygosity. A dual colour FISH showed that in all accessions, the satellite chromosomes carrying the 18S-25S loci did not carry the 5S loci. The results demonstrate that molecular cytogenetics can be applied to Musa and that physical cytogenetic maps can be generated. This revised version was published online in July 2006 with corrections to the Cover Date.     

Growth of arbuscular mycorrhizal mycelium in calcareous dune sand and its interaction with other soil microorganisms as estimated by measurement of specific fatty acids

Fatty acid analysis was used for determining the extent of the development of the external mycelium of AM fungi (mixed inoculum from a sand dune) growing from roots of Festuca rubra and Plantago lanceolata in calcareous dune sand. The plants were raised in chambers specially designed to permit the growth of AM mycelium in root-free compartments. In two separate experiments, growth of external mycelium in the root-free compartments was detected and the amount of mycelium was estimated using the indicator of AM fungal biomass, phospholipid fatty acid (PLFA) 16:15. The results showed that the PLFA 16:15 was suitable for estimating the mycelium emerging from the mixed inoculum obtained from the field roots of F. rubra and P. lanceolata.The PLFA 16:15 showed external mycelium to become established in the root-free compartments within a period of 3 weeks and the amount of mycelium to continue to increase at 6 and 9 weeks. Increases in neutral lipid fatty acid (NLFA) 16:15 (indicator of storage lipids) over time were inconsistent between the two experiments, but appeared to follow patterns of sporulation in each experiment.In both experiments, the root-free compartment was colonised by saprophytic fungi to a greater extent in the case of non-mycorrhizal than of AM treatment, as indicated by an increase in PLFA 18:26,9 (indicator of saprophytic fungi). The absence of an increase in the case of AM treatment indicates that AM fungal mycelium can negatively affect the growth of saprophytic fungi in this soil type. This result was, however, only weakly supported by measurements of ergosterol content. The analysis of bacteria specific PLFAs showed that bacterial biomass was not affected by the AM mycelium.     

水杨酸提高香蕉幼苗的抗冷性与H2O2代谢有关

探讨了水杨酸(salicylic acid, SA)提高香蕉幼苗抗冷性的可能机理.在常温下(30/22 ℃)用不同浓度(0～3.5 mmol/L)的SA水溶液喷洒叶片1 d,置于7 ℃低温下冷胁迫3 d,随后于常温下恢复2 d后测定电解质泄漏率,结果表明:SA 0.3～0.9 mmol/L能显著提高香蕉幼苗的抗冷性,以0.5 mmol/L效果最佳.若把冷胁迫温度降到5 ℃,SA 0.5 mmol/L 预处理可显著减少幼苗叶片的萎蔫面积.但当SA浓度高于1.5 mmol/L时,恢复期间的电解质泄漏甚至高于对照(蒸馏水处理),表明它们加剧了冷害.SA提高香蕉幼苗的抗冷性可能需要H2O2的参与:1)SA 0.5 mmol/L常温处理诱导了H2O2的积累和活性氧造成的膜脂过氧化--三氯乙酸反应物质(TBARS)的增加,这可能与H2O2的清除酶--过氧化氢酶(CAT)和抗坏血酸过氧化物酶(APX)活性的受抑和H2O2的产生酶-超氧化物歧化酶(SOD)活性几乎不受影响有关;2)外源H2O2(1.5～2.5 mmol/L)也能显著降低低温胁迫期间的电解质泄漏,表明也能提高抗冷性;3)而用H2O2的捕捉剂--二甲基硫脲(DMTU)可明显抑制SA诱导的抗冷性;4)在低温胁迫与恢复期间,SA预处理明显提高了CAT和APX的活性,抑制了H2O2与TBARS的快速上升.     

Induction and recovery of copy number variation in banana through gamma irradiation and low‐coverage whole‐genome sequencing

Traditional breeding methods are hindered in bananas due to the fact that major cultivars are sterile, parthenocarpic, triploid and thus clonally propagated. This has resulted in a narrow genetic base and limited resilience to biotic and abiotic stresses. Mutagenesis of in vitro propagated bananas is one method to introduce novel alleles and broaden genetic diversity. We previously established a method for the induction and recovery of single nucleotide mutations generated with the chemical mutagen EMS. However, officially released mutant banana varieties have been created using gamma rays, a mutagen that can produce large genomic insertions and deletions (indels). Such dosage mutations may be important for generating observable phenotypes in polyploids. In this study, we establish a low‐coverage whole‐genome sequencing approach in triploid bananas to recover large genomic indels caused by treatment with gamma irradiation. We first evaluated the commercially released mutant cultivar ‘Novaria’ and found that it harbours multiple predicted deletions, ranging from 0.3 to 3.8 million base pairs (Mbp). In total, predicted deletions span 189 coding regions. To evaluate the feasibility of generating and maintaining new mutations, we developed a pipeline for mutagenesis and screening for copy number variation in Cavendish bananas using the cultivar ‘Williams’. Putative mutations were recovered in 70% of lines treated with 20 Gy and 60% of the lines treated with 40 Gy. While deletion events predominate, insertions were identified in 20 Gy‐treated material. Based on these results, we believe this approach can be scaled up to support large breeding projects.     

落葵的花粉粒和胚囊的形成与发育

落葵(Basella rubra L.)小孢子的发育为同时型,四分体呈四面体排列。在花粉母细胞减数分裂过程中,可清楚地看到胼胝壁的消长变化。三胞花粉,具散沟和网状纹饰。腺质绒毡层。 胚囊的发育为蓼型。反足细胞早期退化。内珠被突出形成珠被喙,同时,在其珠孔区域,有由内珠被的内表皮细胞所发育而来的类似盖状结构。 本文还注意到同一花中以及相邻花朵之间,花粉粒与胚囊发育进程的相关。花粉粒的发育略早于胚囊的发育,但以后由于胚囊母细胞形成胚囊的进程较为迅速,以致花粉粒与胚囊能够同时成熟。     

Antibacterial Δ1‐3‐Ketosteroids from the South China Sea Gorgonian Coral Subergorgia rubra

Three new Δ¹‐3‐ketosteroids characterized with a 9‐OH, subergosterones A–C ( 1 – 3 ), together with five known analogs 4 – 8 , were obtained from the gorgonian coral Subergorgia rubra collected from the South China Sea. The structures of 1 – 3 , including their absolute configurations, were determined by comprehensive spectroscopic methods and electronic circular dichroism (ECD) experiments. Compounds 2 and 3 exhibited inhibitory antibacterial activities against Bacillus cereus with MIC values of 1.56 μM .     

Plantlet production from the male inflorescence tips of <Emphasis Type="Italic">Musa acuminata</Emphasis> cultivars from South India

Inflorescence apices are suitable explants for the rapid in vitro propagation of Musa spp. However, the diploid and triploid banana cultivars showed different in vitro responses with respect to the hormone combinations in Murashige and Skoog medium. The diploid cultivar (Sannachenkadali, AA) induced a maximum number of multiple shoots in 8.9 μM 6-benzyl adenine (BA) whereas the triploid cultivar (Red banana, AAA) exhibited maximum multiplication in 22.2 μM 6-benzyl adenine. MS medium supplemented with 11.4 μM indole acetic acid and 17.8 μM BA was also suitable for shoot proliferation in triploid cultivar but not in the diploid cultivar. The regenerated shoots were rooted in Murashige and Skoog basal medium within 10–15 days. The rooted plantlets were transferred to vermiculite and maintained at a temperature of 25 ± 2°C for 10 days and then at room temperature (30–32°C) for 2 weeks before transferring to potted soil compost mixture. The plantlets showed 100% survival.     

Constituents of Musa balbisiana seeds and their activity against Cryptolestes pusillus

The chloroform and acetone extracts of the seeds of Musa balbisiana Colla (Musaceae) and four of its constituents were assayed in the diet for toxicity against the flat grain beetle (Cryptolestes pusillus Schöherr). A mixture of fatty esters of phytol and the flavan (+)-epiafzelechin showed a significant growth inhibition and toxicity against this economically important pest of stored cereals. The LD₅₀ was 6.3% for the most active compound, (+)-epiafzelechin.     

Nutrient dynamics and ecosystem metabolism in the Bay of Blanes (NW Mediterranean)

The dynamics of the nutrient pools and their stoichiometry as well as their control by ecosystem metabolism (benthic and planktonic) and benthic–pelagic exchanges (sedimentation rates and sediment waterfluxes) were examined in the Mediterranean littoral (Blanes Bay, NE Spain). Dissolved organic nitrogen comprised about half of the nitrogen present in the water column and the carbon pool was dominated by the inorganic pool (95% of the carbon present in the water column). The dissolved and particulate organic pools were deficient in P relative to C and N, indicating a rapid recycling of P from organic matter. The pelagic compartment was heterotrophic, supported by significant allochthonous inputs of land material, which also contributed greatly to the sedimentary inputs (37% of total sedimenting carbon). In contrast, the benthic compartment was autotrophic, with the excess net benthic community production balancing the deficit in pelagic community production, leading to metabolic equilibrium at the station studied. Sedimentary inputs of nitrogen, phosphorus and silicon exceeded the benthic release, indicating that the benthic compartment acted as a sink for nutrients, consistent with its autotrophic nature. Carbon inputs to the benthic compartment also exceeded requirements, due to the allochthonous subsidies to the system, so that the benthic compartment stored or exported organic carbon. An erratum to this article can be found at .     

Purification of an isoflavonoid 7-O-beta-apiosyl-glucoside beta-glycosidase and its substrates from Dalbergia nigrescens Kurz

A beta-glycosidase was purified from the seeds of Dalbergia nigescens Kurz based on its ability to hydrolyse p-nitrophenyl beta-glucoside and beta-fucoside. This enzyme did not hydrolyze various glycosidic substrates efficiently, so it was used to identify its own natural substrates. Two substrates were identified, isolated and their structures determined as: compound 1, dalpatein 7-O-beta-D-apiofuranosyl-(1-->6)-beta-D-glucopyranoside and compound 2, 6,2',4',5'-tetramethoxy-7-hydroxy-7-O-beta-D-apiofuranosyl-(1-->6)-beta-D-glucopyranoside (dalnigrein7-O-beta-D-apiofuranosyl-(1-->6)-beta-D-glucopyranoside). The beta-glycosidase removes the sugar from these glycosides as a disaccharide, despite its initial identification as a beta-glucosidase and beta-fucosidase.