Genetic Diversity in Musa acuminata Colla and Musa balbisiana Colla and some of their natural hybrids using AFLP Markers

Genetic diversity and relationships were assessed in 28 accessions of Musa acuminata (AA) Colla and Musa balbisiana (BB) Colla, and some of their natural hybrids, using the amplified fragment length polymorphisms (AFLP) technique. Fifteen AFLP +3 primer pairs produced 527 polymorphic bands among the accessions. Neighbor-joining and principal co-ordinate (PCO) analyses using Jaccard's similarity coefficient produced four major clusters that closely corresponded with the genome composition of the accessions (AA, BB, AAB and ABB). The AFLP data distinguished between the wild diploid accessions and suggested new subspecies relationships in the M. acuminata complex that are different from those based on morphological data. The data suggested that there are three subspecies within the M. acuminata complex (ssp. burmannica Simmonds, malaccensis Simmonds, and microcarpa Simmonds). 'Tjau Lagada' (ssp. microcarpa), 'Truncata' [ssp truncata (Ridl.) Shepherd] and 'SF247' [ssp. banksii (F.Muell) Simmonds] clustered very closely with 'Gros Michel' and 'Km 5', indicating that more than one M. acuminata subspecies may be involved in the origin of triploid AAA bananas. 'Calcutta 4' (ssp. burmannicoides De Langhe &; Devreux) and 'Long Tavoy' (ssp. burmannica) were closely related and could be together in the same subspecies. This study also showed that there is much more genetic diversity within M. balbisiana that was split into two groups: (1) 'I-63' and 'HND' and (2) 'Los Banos', 'MPL' (Montpellier), '10852', 'Singapuri', 'Etikehel', and 'Butohan 1' as the other.     

Genetic diversity and species-specific PCR-based markers from AFLP analyses of Thai bananas

A large amount of banana genetic resource has been found in Thailand which is believed to be one of the centers of its origins. To assess genetic diversity and determine genetic relationships of edible bananas in Thailand, 110 accessions of banana species and cultivars collected from villages and natural locations were investigated. UPGMA clustering of numerical data from Amplified Fragment Length Polymorphism (AFLP) patterns showed two large groups which corresponded to genome designations of Musa acuminata (AA) and Musa balbisiana (BB), the known ancestors of most edible cultivars. The AFLP data suggested that among Thai bananas, AA and AAA cultivars were closely related to M. acuminata subsp. malaccensis, while some of ‘B’ genome contained ones closely related to wild M. balbisiana in Thailand and some may have been imported. Eight species-specific PCR-based primer pairs, generated from the AFLP results clearly identify ‘A’ and ‘B’ genomes within cultivars and hybrids. The analyses were useful to readily and easily infer progenitors of these cultivars and pronounce wide genetic diversity of the bananas in Thailand.     

First record of the emesine assassin bug genus Emesopsis (Heteroptera: Reduviidae) from Vietnam, with descriptions of two new species

The emesine assassin bug genus Emesopsis is reported from Vietnam for the first time and is briefly diagnosed, and two new species of the genus, E. longipilosa and E. albispinosa, are described. They were collected by beating dead, drooping leaves of the banana, Musa acuminata (Musaceae).     

Study of Resistance of Musa acuminata to Fusarium oxysporum using RAPD markers

Suckers collected from different populations of Musa acuminata ssp. malaccensis were found to be highly resistant to race 4 of Fusarium oxysporum f. sp. cubense (FOC) suggesting that local wild banana populations co-evolved with the pathogen. Seedlings from these wild banana plants segregated for resistance to the pathogen. The infected seedlings were characterized based on external and internal symptoms and the variable response to FOC was mainly due to the genetic factors. Using the technique of random amplified polymorphic DNA (RAPD), 96 major amplification products from 15 primers were identified. Only 10 out of 96 markers were monomorphic and shared among the seed progenies, whereas the remaining 86 were highly polymorphic. Three primers showed banding patterns specific to resistant or susceptible seedlings. These results showed the great potential of the wild Musa acuminata ssp. malaccensis as a source for banana improvement and also for the synthesis of segregating populations for linkage mapping, gene cloning and DNA markers related to FOC resistance.     

Distribution of Lateral Root Primordia in Root Tips of Musa acuminata Colla

The distribution of lateral root primordia in Musa acuminatashows discrete elements of pattern, a major element of whichis the rather regular spacing of laterals along protoxylem-basedranks. There is some co-ordination of positions of lateralsin different ranks. Laterals are apparently not initiated ina single acropetal sequence within the root tip as a whole althoughthey are initiated in acropetal sequence within each rank. Musa acuminata, banana, roots, lateral roots     

Somatic embryogenesis and plant regeneration through cell suspensions inMusa acuminata

Summary Cell suspensions ofMusa acuminata sspburmannicoides andMusa acuminata sspmalaccensis were obtained by culturing embryogenic callus initiated from immature zygotic embryos in liquid medium. Plant regeneration was then achieved through somatic embryogenesis. Germination of these embryos occurred in a modified MS medium containing auxin and cytokinin. Plant recovery frequencies were 20 to 36%. This method may allow a better utilization of biotechnologies in genetic improvement of theMusa diploid species, essential for banana and plantain breeding.     

Relationship Between Lateral Root Primordia in Different Ranks

Comparisons with theoretical random distributions indicate thatthe longitudinal positional relationships between lateral rootsin different ranks are random in roots of Musa acuminata, Pistiastratiotes and i, species in which lateral root primordia arisein the root tip. In Potentilla palustris, where lateral rootprimordia arise further back in the root, there is a deficiencyof close spacings, which indicates that there is some interactionbetween ranks. Musa acuminata, Pistia stratiotes, Pontederia cordat, Potentilla palustris, roots, lateral roots, pattern     

Genome ancestry mosaics reveal multiple and cryptic contributors to cultivated banana

Hybridizations between closely related species commonly occur in the domestication process of many crops. Banana cultivars are derived from such hybridizations between species and subspecies of the Musa genus that have diverged in various tropical Southeast Asian regions and archipelagos. Among the diploid and triploid hybrids generated, those with seedless parthenocarpic fruits were selected by humans and thereafter dispersed through vegetative propagation. Musa acuminata subspecies contribute to most of these cultivars. We analyzed sequence data from 14 M. acuminata wild accessions and 10 M. acuminata‐based cultivars, including diploids and one triploid, to characterize the ancestral origins along their chromosomes. We used multivariate analysis and single nucleotide polymorphism clustering and identified five ancestral groups as contributors to these cultivars. Four of these corresponded to known M. acuminata subspecies. A fifth group, found only in cultivars, was defined based on the ‘Pisang Madu’ cultivar and represented two uncharacterized genetic pools. Diverse ancestral contributions along cultivar chromosomes were found, resulting in mosaics with at least three and up to five ancestries. The commercially important triploid Cavendish banana cultivar had contributions from at least one of the uncharacterized genetic pools and three known M. acuminata subspecies. Our results highlighted that cultivated banana origins are more complex than expected – involving multiple hybridization steps – and also that major wild banana ancestors have yet to be identified. This study revealed the extent to which admixture has framed the evolution and domestication of a crop plant.     

Analysis of genetic diversity and sectional relationships in Musa using AFLP markers

The AFLP technique was used to assess the genetic diversity and sectional relationships in 39 accessions representing the four main sections of the genus Musa. Eight AFLP + 3 primer pairs produced 260 polymorphic bands that were used in cluster and PCO analysis. A wide range of variability was observed among the species within the sections of the genus Musa. AFLP data was useful in separating the different sections of the genus as well as differentiating the different genomic groups of section Eumusa. Section Rhodochlamys (x = 11) appeared as a distinct entity and clustered closely with the Musa acuminata Colla complex of section Eumusa that has the same basic chromosome number. This relationship is congruent with previous studies. However, unlike previous proposals that questioned the identity of Rhodochlamys as a separate taxonomic unit, PCO analysis of the AFLP data showed that it is a distinct entity. Musa laterita Cheesman (Rhodochlamys) and Musa schizocarpa Simmonds clustered with the M. acuminata complex suggesting that they may be sources of useful genes for the improvement of the cultivated bananas. Callimusa formed a distinct unit and was closer to Australimusa than to the other sections. Although both sections share the same basic chromosome number of x = 10 these sections are genetically distinct     

Phylogenetic analysis and differentiation of Veronica subgenus Stenocarpon in the Balkan Peninsula

The Balkan Peninsula is considered the most important refugium for species during the Pleistocene glaciations and today harbours c. 2000 endemic species, but we know surprisingly little about the evolution of taxa in this region. Veronica saturejoides, V. thessalica and V. erinoides are a group of closely related alpine taxa endemic to the Balkan Peninsula. Here, we analyse four DNA regions [the nuclear chalcone synthase intron (CHSi) and ribosomal internal transcribed spacer (ITS) region and the plastid rpoB‐trnC spacer and trnL‐trnL‐trnF region] and amplified fragment length polymorphism (AFLP) fingerprints to provide a phylogenetic hypothesis for the relationships among these taxa. Additionally, we analyse leaf morphological characters used to distinguish the three subspecies of V. saturejoides. The analyses support the distinction of the three subspecies based on previously intuitively suggested characters. Nuclear chalcone synthase intron data indicate that the southern taxa are genetically much more diverse than the more northern V. saturejoides subsp. saturejoides. Phylogenetic relationships inferred from this region and AFLP fingerprints support the monophyly of V. saturejoides. In contrast, plastid DNA regions suggest a closer relationship of V. saturejoides subsp. saturejoides to V. thessalica. The most likely scenario involves introgression into V. saturejoides subsp. saturejoides from V. thessalica. © 2009 The Linnean Society of London, Botanical Journal of the Linnean Society, 2009, 159 , 616–636.     

Sectional relationships in the genus <Emphasis Type="Italic">Musa</Emphasis> L. inferred from the PCR-RFLP of organelle DNA sequences

The objective of this study was to construct a molecular phylogeny of the genus Musa using restriction-site polymorphisms of the chloroplast (cpDNA) and mitochondrial DNA (mtDNA). Six cpDNA and two mtDNA sequences were amplified individually in polymerase chain reaction (PCR) experiments in 13 species representing the four sections of Musa. Ensete ventricosum (W.) Ch. was used as the outgroup. The amplified products were digested with ten restriction endonucleases. A total of 79 restriction-site changes were scored in the sample. Wagner parsimony using the branch and bound option defined two lines of evolution in Musa. One lineage comprised species of the sections Australimusa and Callimusa which have a basic number of x = 10 chromosomes, while most species of sections Eumusa and Rhodochlamys (x = 11) formed the other lineage. Musa laterita Cheesman (Rhodochlamys) had identical organellar genome patterns as some subspecies of the Musa acuminata Colla complex. The progenitors of the cultivated bananas, M. acuminata and Musa balbisiana Colla, were evolutionarily distinct from each other. Musa balbisiana occupied a basal position in the cladogram indicating an evolutionarily primitive status. The close phylogenetic relationship between M. laterita and M. acuminata suggests that species of the section Rhodochlamys may constitute a secondary genepool for the improvement of cultivated bananas.Communicated by H.F. Linskens     

Infraspecific differentiation in the Balkan diploidCardamine acris (Brassicaceae): Molecular and morphological evidence

Molecular and morphological variation in the Balkan diploidCardamine acris was studied by amplified fragment length polymorphism (AFLP) and multivariate morphometric analyses. Principal coordinate analysis and neighbour-joining analysis of the AFLP data showed three genetically differentiated groups of populations corresponding to their geographic origin. Following the results of molecular analysis three subspecies were recognized by a combination of morphological characters as shown by separate analyses of field-collected and cultivated plants. These subspecies are allopatric;C. acris subsp.acris occurs in most parts of the Balkan Peninsula, and newly described subsp.pindicola and subsp.vardousiae are endemic to the Northern Pindhos Mts. in Northwestern Greece, and Central Greece, respectively. The differentiation into geographically isolated subspecies is most probably caused by long-term isolation of populations in different mountain ranges of the Balkan Peninsula, possibly initiated during the cold periods of the Pleistocene.     

Responses of some tropical and subtropical cultures to endomycorrhizal fungi

This paper reports the effects under green-house and field conditions of four arbuscular-mycorrhizal (AM) fungi on the growth and nutrition of some tropical and subtropical crops of economic importance in the Canary Islands (Spain) whose mycorrhizal dependency was unknown. Avocado (Persea americana Mill.), papaya (Carica papaya L.), pineapple (Ananas comosus (Merr.) L.) and banana (Musa acuminata Colla) were highly responsive to the mycorrhizal condition when the AM inoculum was Glomus sp. endophytes. Under our experimental conditions, Glomus fasciculatum was the most effective fungus at improving plantlet growth and nutrition. Acaulospora sp. was ineffective and Scutellospora was only effective with banana. The data clearly show the advantage of mycorrhization during the first phase of development of the plant system.     

Gc subtyping in Malaysians and in Indonesians from North Sumatra

Summary Malays, Chinese and Indians from peninsular Malaysia; Ibans and Bidayuh from Sarawak state, Northern Borneo; and Bataks, Minangkabau and Javanese from North Sumatra, Indonesia, were subtyped for Gc (group-specific component) by polyacrylamide gel isoelectric focusing. All eight populations investigated were found to be polymorphic for three common alleles, Gc^IF, Gc^IS and Gc².     

Using seminatural and simulated habitats for seed germination ecology of banana wild relatives

1. Ecologically meaningful seed germination experiments are constrained by access to seeds and relevant environments for testing at the same time. This is particularly the case when research is carried out far from the native area of the studied species.
2. Here, we demonstrate an alternative—the use of glasshouses in botanic gardens as simulated‐natural habitats to extend the ecological interpretation of germination studies. Our focal taxa were banana crop wild relatives (Musa acuminata subsp. burmannica, Musa acuminata subsp. siamea, and Musa balbisiana), native to tropical and subtropical South‐East Asia. Tests were carried out in Belgium, where we performed germination tests in relation to foliage‐shading/exposure to solar radiation and seed burial depth, as well as seed survival and dormancy release in the soil. We calibrated the interpretation of these studies by also conducting an experiment in a seminatural habitat in a species native range (M. balbisiana—Los Baños, the Philippines), where we tested germination responses to exposure to sun/shade. Using temperature data loggers, we determined temperature dynamics suitable for germination in both these settings.
3. In these seminatural and simulated‐natural habitats, seeds germinated in response to exposure to direct solar radiation. Seed burial depth had a significant but marginal effect by comparison, even when seeds were buried to 7 cm in the soil. Temperatures at sun‐exposed compared with shaded environments differed by only a few degrees Celsius. Maximum temperature of the period prior to germination was the most significant contributor to germination responses and germination increased linearly above a threshold of 23℃ to the maximum temperature in the soil (in simulated‐natural habitats) of 35℃.
4. Glasshouses can provide useful environments to aid interpretation of seed germination responses to environmental niches.

     

Fossil bananas (Musaceae): Ensete oregonense sp. nov. from the Eocene of western North America and its phytogeographic significance

Fossil seeds of Ensete, a genus presently native to Asia and Africa, have been recovered from the middle Eocene of Oregon, confirming the presence of Musaceae in the North American Tertiary. The seed of Ensete oregonense sp. nov. is operculate, with a well-defined micropylar collar, a pronounced chalazal chamber, and a wide hilar cavity. A survey of seed morphology in extant Zingiberales provides characters for distinguishing Musaceae from other families of the order, furnishes criteria for distinguishing the three extant genera of Musaceae (Musa, Ensete and Musella), and facilitates critical assessment of fossil seed remains. “Musa” cardiosperma Jain from the Cretaceous/Tertiary boundary Deccan Series of India is excluded from Musaceae (although retained in Zingiberales) on the basis of fruit and seed characters, including the lack of laticifers and absence of a chalazal chamber. We reexamined the musaceous seeds from Colombia that previously were described as Tertiary fossils (Musa enseteformis Berry, 1925) and now believe that they are recent, nonfossil remains, evidently from Ensete ventricosum, which is grown in the region where the specimens were originally obtained. In addition, a reputed fossil banana fruit from the Cretaceous of Colombia was reexamined and determined to be a concretion of nonbiological origin. Ensete oregonense is significant therefore, as the first unequivocal fossil record of Ensete and of Musaceae. Although the Musaceae are currently native only to the Old World tropics, this discovery establishes that the family was present in North America about 43 million years ago.     

Taxonomic revision of shrews of the genus Crociduras from the Sunda Shelf and Sulawesi with description of two new species (Mammalia: Soricidae)

The skulls of 387 shrews of the genus Crocidura sampled in peninsular Malaysia, Sumatra, Borneo, Java and Sulawesi were submitted to principal component and stepwise discriminant analyses. These analyses helped to delineate morphological taxa in this species-rich genus of mammals. Most morphologic groups could be attributed to described species, except one taxon from Sumatra and one from Sulawesi, which are described and named as new. Most of the 21 species recognized in this paper are endemic to one major island. Although Sulawesi has never been connected to the mainland, it supports at least six species, followed by Sumatra (5–6 species), Java and the Malay Peninsula (4 spp) and Borneo (3 spp). C. monticola is apparently the only widespread species whose distribution range covers the entire Malay Archipelago except the Philippines and Sulawesi. In contrast, the continental C. fuliginosa enters only marginally into the Sunda Shelf: its southernmost record is on the Malay Peninsula. This interpretation is completely different from the classical view that C. fuliginosa is a cosmopolitan species occupying the whole of Southeast Asia. Identification keys, tables of measurements and discriminant functions provided in this work may aid in identification of the various species and subspecies of Crocidura living in the Malaya Archipelago.     

Characterization of disease resistance gene candidates of the nucleotide binding site (NBS) type from banana and correlation of a transcriptional polymorphism with resistance to <Emphasis Type="Italic">Fusarium oxysporum</Emphasis> f.sp. <Emphasis Type="Italic">cubense</Emphasis> race 4

Most plant disease resistance (R) genes encode proteins with a nucleotide binding site and leucine-rich repeat structure (NBS-LRR). In this study, degenerate primers were used to amplify genomic NBS-type sequences from wild banana (Musa acuminata ssp. malaccensis) plants resistant to the fungal pathogen Fusarium oxysporum formae specialis (f. sp.) cubense (FOC) race 4. Five different classes of NBS-type sequences were identified and designated as resistance gene candidates (RGCs). The deduced amino acid sequences of the RGCs revealed the presence of motifs characteristic of the majority of known plant NBS-LRR resistance genes. Structural and phylogenetic analyses grouped the banana RGCs within the non-TIR (homology to Toll/interleukin-1 receptors) subclass of NBS sequences. Southern hybridization showed that each banana RGC is present in low copy number. The expression of the RGCs was assessed by RT-PCR in leaf and root tissues of plants resistant or susceptible to FOC race 4. RGC1, 3 and 5 showed a constitutive expression profile in both resistant and susceptible plants whereas no expression was detected for RGC4. Interestingly, RGC2 expression was found to be associated only to FOC race 4 resistant lines. This finding could assist in the identification of a FOC race 4 resistance gene.     

Four new species, a new variety, and a status change in Argostemma (Rubiaceae) from Thailand

Four new species of Argostemma , all endemic to Thailand are described: A. thaithongae , restricted to northern Thailand; A. monophyllum , rather widely distributed in the easern half of Thailand: A. puffii and A. routundicalyx , both endemic to peninsular Thailand. Var. variabile is a new variety of the peninsular Thailand, species A. lobulatum. A. setosum , previously thought to be an endemic species of peninsular Thailand, is now considered to be a subspecies of A. laeve (Malay peninsula and Sumatra): A. laeve subsp. setosum , stat. nov.