Archaeological manioc (Manihot) from Coastal Peru

Preserved remains of manioc(Manihot) from 6 archaeological sites in the Casma Valley of Peru are illustrated and described. The combined collections from these sites total 197 pieces of root, 32 bark fragments, 22 pieces of stem, 4 capsules, and 2 leaf twigs. Based on radiocarbon assays, the specimens range in age from 1800 B.C. to A.D. 1532. This collection of sweet manioc is unique for its age, number, and wealth of different plant parts. A theory on the place of origin and time of domestication of these ancient cultivars is given in the conclusions of this paper.     

Starch granule size and the domestication of manioc (Manihot esculenta) and sweet potato (Ipomoea batatas)

Archaeological studies of plant remains have indicated that an increase in seed size is frequently correlated with both intensive cultivation and domestication of seed crop plants. To test if starch granules of domesticated root crops are significantly larger than those of wild or less intensively cultivated plants, archaeological and modern specimens of manioc and sweet potato were sampled for starch granules, and granule size was compared across a temporal sequence. The results indicate that a gross generalization can be made that modern specimens of both manioc and sweet potato yield larger starch granules than some archaeological specimens. It does appear, however, that modern domesticated manioc roots produce significantly larger-sized starch granules than those of its purported wild ancestor. Additionally, there exist two lines of evidence that the coastal Peruvian and lowland Neotropical regional types of manioc differ from one another and have been separate for several millennia. These findings indicate that manioc may have been domesticated more than once.     

Cyanide content of cassava (Manihot esculenta,Euphorbiaceae) cultivars used by Tukanoan Indians in Northwest Amazonia

Cassava (Manihot esculenta) is a cyanide-containing food crop used by many indigenous peoples in Amazonia. Tukanoan Indians in Northwest Amazonia utilize both “bitter” and “sweet” cassava cultivars. Those classified as “bitter” are the dietary staple. Analysis of 13 commonly used “bitter” cultivars indicates that they are high in cyanide in comparison to values reportedin the literature. The Tukanoan practice of including the inner peel in the edible portion contributes to the high cyanide values found.     

Isoforms of Linamarase in Cassava (Manihot esculenta)

Linamarase (EC. 3.2.1.21) was purified from different tissues of cassava (leaf, rind and tuber) to compare the kinetic properties and characteristics of the enzyme in these tissues. Purified enzyme preparation appeared as single band of average molecular size 70 kD in SDS-PAGE gels. The kinetic properties of linamarase with respect to pH and temperature indicated that tuber linamarase possessed a broader pH optimum and higher temperature stability as compared to leaf and rind enzymes. Differences in Km values for linamarin were observed with leaf linamarase having the highest Km value (500 μM) followed by rind (400 μM) and then tuber (250 μM) linamarases. Rind enzyme appeared to be less susceptible to urea denaturation than the leaf enzyme. Comparison of elution profiles from DEAE-Cellulose indicated that the relative amounts of the various ionic forms of the enzyme differed in the case of each tissue. Elution patterns of the enzyme from Con A-Sepharose also differed, suggesting difference in glycosylation among leaf, rind and tuber enzymes. This was confirmed by carbohydrate analysis which showed that the tuber linamarase contained significantly higher amount of protein bound carbohydrate. These results suggest the possible occurrence of different forms of linamarase in cassava.     

Biological parameters of Bemisia tabaci (Gennadius) biotype B (Hemiptera: Aleyrodidae) on Jatropha gossypiifolia, commercial (Manihot esculenta) and wild cassava (Manihot flabellifolia and M. carthaginensis) (Euphorbiaceae)

Bemisia tabaci (Gennadius) is one of the most important pests of cassava in Africa and several countries of Asia due to the damage caused by direct feeding, the excretion of honeydew, and its capacity as a vector of cassava mosaic geminivirus. There is a general consensus that B. tabaci is a complex of morphologically indistinguishable populations with different biotypes. In the Americas, the polyphagous biotype B does not appear to feed on cassava. Recent studies indicate that it is possible, however, for biotype B to gradually adapt to cassava using phylogenetically related hosts. Therefore, the possibility that some wild species of cassava constitute intermediate hosts in the adaptation process may lead to the establishment of biotype B on commercial varieties of Manihot esculenta. In here, we evaluated Jatropha gossypiifolia, two wild species of cassava (Manihot flabellifolia and M. carthaginensis) and a commercial cassava variety (MCol 2063) as hosts of biotype B. The highest oviposition rate (2.7 eggs /two days) occurred on M. esculenta, although the development time (44 d) was the longest when compared to M. carthaginensis and J. gossypiifolia. About 60% of the population could reproduce on the wild cassava species vs. 55% on J. gossypiifolia and 27.5% on the commercial variety. Our data suggest that J. gossypiifolia is a suitable host and the wild species M. carthaginensis can constitute a potential intermediate host in the adaptation of biotype B to commercial varieties of cassava.     

In vitro propagation of cassava (Manihot esculenta Crantz)

A method is presented for the rapid in vitro propagation of cassava (Manihot esculenta Crantz). Nodal explants were induced to grow as multiple-shoot cultures on a medium containing 1.0 M 6-benzylamino purine (BAP), supplemented with 0.25 M -naphthaleneacetic acid (NAA). Nodes were removed from the shoots after three weeks of growth and subcultured on fresh culture medium. An average of 7.0 nodes were produced from each explanted node after three weeks in culture. Nodal explants were transferred to a medium containing 2.5 M indole-3-butyric acid (IBA) to improve root initiation on the developing plantlets. Plant establishment was possible upon transfer to soil. In vitro propagation offers enhanced rates of multiplication over more conventional methods of propagation. In addition, in vitro propagation facilitates the storage and international exchange of cassava germplasm.     

In vitro flowering in cassava (Manihot esculenta Crantz)

Meristem-derived plantlets of cassava (Manihot esculenta Crantz) were induced to flower in vitro. Five genotypes out of 13 consistently responded to our culture conditions giving rise to male or female flowers. Male flowers contained anthers in which meiosis occurred and apparently normal pollen grains were formed.     

Isozyme Diversity in Cassava Cultivars (Manihot esculenta Crantz)

Isoenzyme electrophoresis was used as a method to determine genetic diversity in various M. esculenta cultivars collected in the Southwestern (SW) and Northwestern (NW) regions of the State of Parana, in the South region of Brazil, and in cultivars produced at the Agronomic Institute of Campinas (IAC), S~ao Paulo State, Southeastern region of Brazil. The cultivars have been maintained by vegetative propagation for 5 years and are useful in production programs. A total of 28 loci in the acid phosphatase (ACP; EC 3.1.3.2), esterases (EST; EC 3.1.1.1), malate dehydrogenase (MDH; EC 1.1.1.37), and shikimate dehydrogenase (SKDH; EC 1.1.1.15) isozymes was analyzed. The proportion of polymorphic loci for NW, SW, and IAC cultivars was 57.14, 50.0, and 53.6%, respectively. Genetic diversity calculated by Nei's genetic identity (I) showed high I values for the three M. esculenta subpopulations. The high degree of polymorphism expressed by cassava cultivars is highly relevant to stimulate breeding programs with M. esculenta species.     

Germination ecology of Cassava (Manihot esculenta Crantz, Euphorbiaceae) in traditional agroecosystems: Seed and seedling biology of a vegetatively propagated domesticated plant

Cassava is clonally propagated, but Amerindian farmers also use plants from volunteer seedlings to prepare stem cuttings. Although sexual reproduction plays a role in cassava’s evolution it is poorly studied. We examined one aspect of cassava reproductive ecology, seed dormancy and germination. Volunteer seedlings emerge from a soil bank of seeds produced during the previous cycle of cultivation that remain ungerminated through the fallow period, then germinate synchronously after vegetation is slashed and burned. Laboratory experiments showed that germination can be enhanced by mechanical scarification and also by dry heat treatment, suggesting that burning after field clearing could help break dormancy. Germination was also stimulated by high temperatures (35°C) that in nature indicate bare soils, and inhibited by temperatures (25°C) close to those in soil shaded by vegetation and by light. Seeds of both wild and domesticated cassava exhibit physiological dormancy, an adaptation for germination in periodically disturbed habitats. In addition to these preadaptations, preliminary results also suggest specific adaptations of domesticated cassava to the distinctive disturbance regimes of swidden agriculture.     

A molecular genetic map of cassava (Manihot esculenta Crantz)

 A genetic linkage map of cassava has been constructed with 132 RFLPs, 30 RAPDs, 3 microsatellites, and 3 isoenzyme markers segregating from the heterozygous female parent of an intraspecific cross. The F₁ cross was made between ‘TMS 30572’ and ‘CM 2177-2’, elite cassava cultivars from Nigeria and Colombia, respectively. The map consists of 20 linkage groups spanning 931.6 cM or an estimated 60% of the cassava genome. Average marker density is 1 per 7.9 cM. Since the mapping population is an F₁ cross between heterozygous parents, with unique alleles segregating from either parent, a second map was constructed from the segregation of 107 RFLPs, 50 RAPDs, 1 microsatellite, and 1 isoenzyme marker from the male parent. Comparison of intervals in the male-and female-derived maps, bounded by markers heterozygous in both parents, revealed significantly less meiotic recombination in the gametes of the female than in the male parent. Six pairs of duplicated loci were detected by low-copy genomic and cDNA sequences used as probes. Efforts are underway to saturate the cassava map with additional markers, to join the male- and female-derived maps, and to elucidate genome organization in cassava. Received: 5 July 1996/Accepted: 22 November 1996     

Characterization of cyanogenic beta-glucosidase (linamarase) from cassava (Manihot esculenta Crantz)

Linamarase (EC 3.2.1.21) was purified from cassava petiole, stem, and root cortex by ammonium sulfate precipitation, column chromatography on Sepharose 6B, and chromatofocusing. The last step resolved the enzyme from each source into three forms with pI values of 4.3, 3.3, and 2.9. Each form was found to be oligomeric, consisting of one kind of subunit, Mr 63,000. The major isozyme with a pI of 4.3 from petiole showed a Km for linamarin of 0.6 mM and possessed both beta-glucosidase and beta-fucosidase activities. The former was sensitive to inhibition by delta-gluconolactone, isopropyl-beta-D-thioglucoside, and HgCl2, whereas the latter was inhibited by Tris ion.     

REVIEW ARTICLE: Cyanogenesis in cassava (Manihot esculenta Crantz)

Cassava is the most agronomically important of the cyanogeniccrops. Linamarin, the predominant cyanogenic glycoside in cassava,can accumulate to concentrations as high as 500 mg kg^–1fresh weight in roots and to higher levels in leaves. Recently,the pathway of linamarin synthesis and the cellular site oflinamarin storage have been determined. In addition, the cyanogenicenzymes, linamarase and hydroxynitrile lyase, have been characterizedand their genes cloned. These results, as well as studies onthe organ- and tissue-specific localization of linamarase andhydroxy-nitrile lyase, allow us to propose models for the regulationof cyanogenesis in cassava. There remain, however, many unansweredquestions regarding the tissue-specific synthesis, transport,and accumulation of cyanogenic glycosides. The resolution ofthe sequestions will facilitate the development of food processing,biochemical and transgenic plant approaches to reducing thecyanogen content of cassava foods. Key words: Cyanide, cyanogenic glycosides, linamarin, cyanogens     

Improvements of cyclic somatic embryogenesis of cassava (Manihot esculenta Crantz)

Summary In cassava a cyclic system of somatic embryogenesis was developed. Primary (torpedo shaped or germinated) embryos, originating from leaf lobes, could only be obtained after culture on solid medium. Cyclic embryos, originating from embryos, could be obtained in both liquid and on solid medium. The production of embryos in liquid medium was distinctly higher, faster and more synchronized than on solid medium. Lower densities and fragmentation of starting embryos improved the production significantly. The highest production found was 32.1 embryos per initial embryo. In all treatments the explants initiated multiple embryos. The production of single embryos was achieved by pressing starting embryos through a fine meshed sieve, indicating that embryos can be produced from a piece of tissue with a restricted number of cells. The shoot conversion rate of embryos from liquid medium was comparable with that of embryos from solid medium.Abbreviations BM Basal Medium - MIE medium volume per initial embryo - E/IE number of Embryos per Initial Embryo     

A geographic distribution database of Mononychellus mites (Acari,Tetranychidae) on cassava (Manihot esculenta)

The genus Mononychellus is represented by 28 herbivorous mites. Some of them are notorious pests of cassava (Manihot esculenta Crantz), a primary food crop in the tropics. With the exception of Mononychellus tanajoa (Bondar), their geographic distribution is not widely known. This article therefore reports observational and specimen-based occurrence data of Mononychellus species associated with cassava. The dataset consists of 1,513 distribution records documented by the International Center for Tropical Agriculture (CIAT) between 1975 and 2012. The specimens are held at CIAT’s Arthropod Reference Collection (CIATARC). Most of the records are from the genus’ native range in South America and were documented between 1980 and 2000. Approximately 61% of the records belong to M. tanajoa, 25% to M. caribbeanae (McGregor), 10% to M. mcgregori (Flechtmann and Baker) and 2% to M. planki (McGregor). The complete dataset is available in Darwin Core Archive format via the Global Biodiversity Information Facility (GBIF).     

The microbial breakdown of linamarin in fermenting pulp of cassava (Manihot esculenta Crantz)

Summary Fifty-five organisms comprising 40 bacteria, eleven yeasts and four moulds were isolated from various habitats and tested for the production of the enzyme linamarase using the release of HCN and glucose from linamarin. Only seven organisms were shown to produce linamarase: the bacteriaLeuconostoc mesenteroides, Alcaligenes faecalis, the yeastsSaccharomyces cerevisiae andRhodotorula minuta, and the mouldsAspergillus flavus, A. niger andFusarium oxysporum. A preliminary screening method used the ability to break down the glucosidep-nitrophenyl--D glucoside (PNPG). Organisms breaking down PNPG also broke down linamarin and vice-versa. The highest linamarase producers among the yeasts and bacteria as determined by HCN liberation from cassava pulp were the yeastsSacch. cerevisiae andR. minuta in about 80 and 96 hours, respectively. SinceSaccharomyces sp. withstood the highest concentration of the cyanide ion it is suggested as the organism of preference among the isolates for cassava pulp detoxication. Storage of linamarase-producing organisms in refrigerated cassava pulp was found to be a suitable method of preserving and transporting them.

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Resumen Cincuenta y cinco organismos, incluyendo cuarenta bacterias, once levaduras y cuatro hongos se aislaron a partir de distintos hábitats y se estudió su producción de enzyma linamarasa, determinada por la liberación de HCN y glucosa a partir de linamarina. Tan solo siete organismos producían linamarasa: las bacteriasLeuconostoc mesenteroides yAlcaligenes faecalis, las levadurasSaccharomyces cerevisiae yRhodotorula minuta y los hongosAspergillus flavus, A. niger yFusarium oxysporum. Como método de estudio preliminar se utilizó la habilidad para degradar el glucósidop-nitrofenil-l-glucósido (PNPG), ya que los organismos capaces de degradar PNPG también eran capaces de degradar linamarina y viceversa. Entre bacterias y levaduras los mayores productores de linamarasa, expresada como cantidad de HCN liberado a partir de pulpa de casava, fueronSaccharomyces cerevisiae yRhodotorula minuta, a las 80 y 96 horas respectivamente. Al soportarSaccharomyces sp. la mayor de las concentraciones de HCN ensayadas se ha sugerido que este podría ser considerado, de entre los organismos aislados, como el más idoneo para ser utilizado en la detoxificación de la pulpa de casava. Se considera que un método adecuado para la preservación de organismos productores de linamarasa es su almacenamiento en pulpa de casava refrigerada.

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Résumé Cinquante-cinq organismes comprenant 40 bactéries, 11 levures et 4 moisissures ont été isolés à partir de différents habitats et testés pour la production de l'enzyme linamarase qui utilise le HCN et le glucose provenant de la linamarine. Une méthode de criblage préliminaire utilise l'aptitude à dégrader lep-nitrophényl-D-glucoside (PNPG). Les organismes qui dégradent le PNPG dégradent aussi la linamarine et réciproquement. Sept organismes seulement produisent la linamarase: les bactériesLeuconostoc mesenteroides etAlcaligenes faecalis, les levuresSaccharomyces cerevisiae etRhodotorula minuta, et les moisissuresAspergillus flavus, A. niger etFusarium oxysporum. D'après la vitesse de libération d'HCN à partir de la pulpe de manioc, les meilleurs producteurs de linamarase sont les levuresS. cerevisiae etR. minuta agissant, respectivement, en 80 et 96 heures environ.Saccharomyces étant parmi les organismes isolés celui qui tolère la concentration en ion cyanure la plus élevée, son utilisation est proposée pour détoxifier la pulpe de manioc. La conservation des organismes producteurs de linamarase dans la pulpe de manioc réfrigérée est la méthode proposée pour la préserrvation et le transport de ces organismes.

     

Anatomic changes due to interspecific grafting in cassava (Manihot esculenta)

Cassava rootstocks of varieties UnB 201 and UnB 122 grafted with scions of Manihot fortalezensis were prepared for anatomic study. The roots were cut, stained with safranin and alcian blue, and examined microscopically, comparing them with sections taken from ungrafted roots. There was a significant decrease in number of pericyclic fibers, vascular vessels and tyloses in rootstocks. They exhibited significant larger vessels. These changes in anatomic structure are a consequence of genetic effects caused by transference of genetic material from scion to rootstock. The same ungrafted species was compared. This is the first report on anatomic changes due to grafting in cassava.     

Response of cassava (Manihot esculenta Crantz) to water stress and fertilization

Experiments done in Santander de Quilichao (Cauca, Colombia) on two cassava cultivars indicated that cassava had at least three defence mechanisms against water deficit, enabling it to assimilate and store photosynthates in roots, even during prolonged droughts. These mechanisms include partial stomatal closure, ability of leaves to maintain reasonable net photosynthetic rate for long periods of water stress, reduced leaf area, and exploration of water from deep soil layers. While cassava responded positively to fertilization, no significant statistical differences were found between treatments of stress and non-stress, confirming cassava's ability to tolerate soil water deficit. This revised version was published online in June 2006 with corrections to the Cover Date.     

Response of cassava (Manihot esculenta Crantz) to water stress and fertilization

Experiments done in Santander de Quilichao (Cauca, Colombia) on two cassava cultivars indicated that cassava had at least three defence mechanisms against water deficit, enabling it to assimilate and store photosynthates in roots, even during prolonged droughts. These mechanisms include partial stomatal closure, ability of leaves to maintain reasonable net photosynthetic rate for long periods of water stress, reduced leaf area, and exploration of water from deep soil layers. While cassava responded positively to fertilization, no significant statistical differences were found between treatments of stress and non-stress, confirming cassava's ability to tolerate soil water deficit.