Analysis of leaf morphology, secondary metabolites and proteins related to the resistance to Tetranychus cinnabarinus in cassava (Manihot esculenta Crantz).

Constitutive resistance of plant can be divided into physical and chemical barriers. Cassava (Manihot esculenta Crantz) is susceptible to mites, especially Tetranychus cinnabarinus. Although significant differences in the resistance to T. cinnabarinus are observed in different cassava cultivars, limited research has been done on the mechanism accounting for the resistance. The aim of this study was to explore the mechanism of resistance to T. cinnabarinus by comparing morphology, secondary metabolites and proteins in different cassava cultivars. The anatomical structure of leaves showed that the cassava cultivar Xinxuan 048 (XX048), which showed a stronger resistance to T. cinnabarinus in both greenhouse testing and three years field evaluation tests (2016-2018), had thicker palisade tissue, spongy tissue, lower epidermis and leaf midrib tissue compared to cultivar Guire 4 (GR4). Greenhouse evaluation demonstrated that originally these cultivars were different, leading to differences in constitutive levels of metabolites. The proteomic analysis of protected leaves in XX048 and GR4 revealed that up-regulated differentially expressed proteins (DEPs) were highly enriched in secondary metabolic pathways, especially in the biosynthesis of flavonoids. This study not only provides a comprehensive data set for overall proteomic changes of leaves in resistant and susceptible cassava, but also sheds light on the morphological characteristics of cassava-mite interaction, secondary metabolite defense responses, and molecular breeding of mite-resistant cassava for effective pest control.

Response of cassava cultivars to African cassava mosaic virus infection across a range of inoculum doses and plant ages.

Cassava production in Africa is constrained by cassava mosaic disease (CMD) that is caused by the Cassava mosaic virus (CMV). The aim of this study was to evaluate the responses of a range of commonly cultivated West African cassava cultivars to varying inoculum doses of African cassava mosaic virus (ACMV). We grafted 10 cultivars of cassava plants with different inoculum doses of CMV (namely two, four, or six CMD-infected buds) when the experimental plants were 8, 10, or 12 weeks old, using non-inoculated plants as controls. Three cultivars showed disease symptoms when grafted with two buds, and four cultivars showed disease symptoms when grafted with four or six buds. Most cultivars became symptomatic six weeks after inoculation, but one ('TMS92/0326') was symptomatic two weeks after inoculation, and two ('Ntollo' and 'Excel') were symptomatic after four weeks. Root weight tended to be lower in the six-bud than in the two-bud dose, and disease severity varied with plant age at inoculation. These results indicate that the level of CMD resistance in cassava cultivars varies with inoculum dose and timing of infection. This will allow appropriate cultivars to be deployed in each production zone of Africa in accordance with the prevalence of CMD.

Patterns of nuclear and chloroplast genetic diversity and structure of manioc along major Brazilian Amazonian rivers.

Background and Aims: Amazonia is a major world centre of plant domestication, but little is known about how the crops were dispersed across the region. Manioc (Manihot esculenta) was domesticated in the south-western Amazon basin, and is the most important staple food crop that originated in Amazonia. Current contrasting distributions may reflect distinct histories of dispersal of bitter and sweet manioc landraces. To produce new insights into the evolutionary history of the crop, we investigated the contemporary genetic diversity and structure of bitter and sweet manioc along major Amazonian rivers. Methods: The patterns of genetic structure and diversity of wild and cultivated sweet and bitter manioc with four chloroplast and 14 nuclear microsatellite markers were evaluated. Results were interpreted in terms of the crop's dispersal. Key results: No phylogeographic patterns among rivers were detected, and genetic structure among rivers was confounded by the bitter-sweet divergence. However, differences in the distribution of nuclear diversity and somewhat distinctive patterns of genetic structure across rivers were observed within bitter and sweet manioc. Conclusions: Various pre-Columbian and post-European conquest events in the history of Amazonian occupation may explain the absence of clearer patterns of genetic structure. However, the wide distribution of the most common chloroplast haplotype agrees with an early dispersal of manioc across Brazilian Amazonia. Furthermore, differences in genetic structure and in the spatial distribution of genetic diversity suggest that bitter and sweet manioc had distinct dispersal histories. Knowledge about how prehistoric and contemporary Amazonian peoples manage their crops is valuable for the maintenance and conservation of the impressive diversity of their native crops.

Leaf area estimation of cassava from linear dimensions

ABSTRACT The objective of this study was to determine predictor models of leaf area of ​​cassava from linear leaf measurements. The experiment was carried out in greenhouse in the municipality of Botucatu, São Paulo state, Brazil. The stem cuttings with 5-7 nodes of the cultivar IAC 576-70 were planted in boxes filled with about 320 liters of soil, keeping soil moisture at field capacity, monitored by puncturing tensiometers. At 80 days after planting, 140 leaves were randomly collected from the top, middle third and base of cassava plants. We evaluated the length and width of the central lobe of leaves, number of lobes and leaf area. The measurements of leaf areas were correlated with the length and width of the central lobe and the number of lobes of the leaves, and adjusted to polynomial and multiple regression models. The linear function that used the length of the central lobe LA = -69.91114 + 15.06462L and linear multiple functions LA = -69.9188 + 15.5102L + 0.0197726K - 0.0768998J or LA = -69.9346 + 15.0106L + 0.188931K - 0.0264323H are suitable models to estimate leaf area of ​​cassava cultivar IAC 576-70.

Morphological diversity of cassava accessions of the south-central mesoregion of the State of Mato Grosso, Brazil.

Genetic variability of cassava (Manihot esculenta Crantz) in Brazil is wide, being this the result of natural and cultural selection during pre- and post-domestication of the species in different environments. Given the number of species of the genus found in the region (38 of a total of 98 species), the central region of Brazil was defined as the primary center of cassava diversity. Therefore, genetic diversity characterization of cassava accessions is fundamental, both for farmers and for plant breeders, because it allows the organization of genetic resources and better utilization of available genetic diversity. This research aims to assess genetic divergence of cassava accessions from the south-central region of the State of Mato Grosso, based on multi-categorical morphological traits. For this purpose, 38 qualitative and quantitative morphological descriptors were used. Genetic diversity was expressed by the genetic similarity index, with subsequent clustering of accessions by the modified Tocher's procedure and UPGMA. Of 38 descriptors, only growth habit of stem showed no variability. Tocher and UPGMA methods were efficient and corroborated on group composition. Both methods were able to group accessions of different localities in distinct group consistency.

Leaf area estimation of cassava from linear dimensions.

The objective of this study was to determine predictor models of leaf area of ​​cassava from linear leaf measurements. The experiment was carried out in greenhouse in the municipality of Botucatu, São Paulo state, Brazil. The stem cuttings with 5-7 nodes of the cultivar IAC 576-70 were planted in boxes filled with about 320 liters of soil, keeping soil moisture at field capacity, monitored by puncturing tensiometers. At 80 days after planting, 140 leaves were randomly collected from the top, middle third and base of cassava plants. We evaluated the length and width of the central lobe of leaves, number of lobes and leaf area. The measurements of leaf areas were correlated with the length and width of the central lobe and the number of lobes of the leaves, and adjusted to polynomial and multiple regression models. The linear function that used the length of the central lobe LA = -69.91114 + 15.06462L and linear multiple functions LA = -69.9188 + 15.5102L + 0.0197726K - 0.0768998J or LA = -69.9346 + 15.0106L + 0.188931K - 0.0264323H are suitable models to estimate leaf area of ​​cassava cultivar IAC 576-70.

Domestication Syndrome Is Investigated by Proteomic Analysis between Cultivated Cassava (Manihot esculenta Crantz) and Its Wild Relatives.

Cassava (Manihot esculenta Crantz) wild relatives remain a largely untapped potential for genetic improvement. However, the domestication syndrome phenomena from wild species to cultivated cassava remain poorly understood. The analysis of leaf anatomy and photosynthetic activity showed significantly different between cassava cultivars SC205, SC8 and wild relative M. esculenta ssp. Flabellifolia (W14). The dry matter, starch and amylose contents in the storage roots of cassava cultivars were significantly more than that in wild species. In order to further reveal the differences in photosynthesis and starch accumulation of cultivars and wild species, the globally differential proteins between cassava SC205, SC8 and W14 were analyzed using 2-DE in combination with MALDI-TOF tandem mass spectrometry. A total of 175 and 304 proteins in leaves and storage roots were identified, respectively. Of these, 122 and 127 common proteins in leaves and storage roots were detected in SC205, SC8 and W14, respectively. There were 11, 2 and 2 unique proteins in leaves, as well as 58, 9 and 12 unique proteins in storage roots for W14, SC205 and SC8, respectively, indicating proteomic changes in leaves and storage roots between cultivated cassava and its wild relatives. These proteins and their differential regulation across plants of contrasting leaf morphology, leaf anatomy pattern and photosynthetic related parameters and starch content could contribute to the footprinting of cassava domestication syndrome. We conclude that these global protein data would be of great value to detect the key gene groups related to cassava selection in the domestication syndrome phenomena.

Comparative petiole anatomy of cassava (Manihot) species.

In this study, we describe the petiole anatomy of six wild cassava (Manihot) species, one hybrid, and two cultivars of Manihot esculenta, in order to identify their dominant anatomical patterns and relate them to possible adaptations to abiotic factors in the Cerrado biome. The median parts of several petiole samples were transversally and longitudinally sectioned and stained. The results include data for the taxonomic classification of the genus, including distinctive anatomical characteristics of hybrid varieties of cassava and wild species, such as the presence/absence of trichomes and a hypodermis, layer type and number in the cortex, number of vascular bundles, cell types in the pith, and type of organization. Morphological analysis revealed differences in length and shape of the petiole insertion. The presence of trichomes, a hypodermis, the amount and type of supporting tissue in the cortex, as well as gelatinous fibers, may be related to drought tolerance.

Cassava root membrane proteome reveals activities during storage root maturation.

Cassava (Manihot esculenta Crantz) is one of the most important crops of Thailand. Its storage roots are used as food, feed, starch production, and be the important source for biofuel and biodegradable plastic production. Despite the importance of cassava storage roots, little is known about the mechanisms involved in their formation. This present study has focused on comparison of the expression profiles of cassava root proteome at various developmental stages using two-dimensional gel electrophoresis and LC-MS/MS. Based on an anatomical study using Toluidine Blue, the secondary growth was confirmed to be essential during the development of cassava storage root. To investigate biochemical processes occurring during storage root maturation, soluble and membrane proteins were isolated from storage roots harvested from 3-, 6-, 9-, and 12-month-old cassava plants. The proteins with differential expression pattern were analysed and identified to be associated with 8 functional groups: protein folding and degradation, energy, metabolism, secondary metabolism, stress response, transport facilitation, cytoskeleton, and unclassified function. The expression profiling of membrane proteins revealed the proteins involved in protein folding and degradation, energy, and cell structure were highly expressed during early stages of development. Integration of these data along with the information available in genome and transcriptome databases is critical to expand knowledge obtained solely from the field of proteomics. Possible role of identified proteins were discussed in relation with the activities during storage root maturation in cassava.

Apparent similarity, underlying homoplasy: Morphology and molecular phylogeny of the North American clade of Manihot.

PREMISE OF THE STUDY: Morphologically diverse clades are useful for detecting adaptive morphological evolution. Each of their variants may have evolved once or several times, suggesting that their repeated appearance may be due to environmental pressures. The North American Manihot species are an excellent system to detect possible adaptations and to assess the effect of mono- or polyphyly on classification. With 20 species, this group includes growth forms from tuberous herbs to trees. The monophyly of this group and its relationship with the economically important M. esculenta were tested for the first time with complete sampling of North American species. METHODS: We carried out maximum likelihood and Bayesian phylogenetic analyses on a matrix of 3662 bp from chloroplast (psbA-trnH, trnL-trnF) and nuclear loci (PEPC and two paralogous copies of G3pdh). We included all North American Manihot species, Manihotoides pauciflora, and published sequences from 34 South American species. KEY RESULTS: Our results support monophyly of the North American Manihot group. Its taxonomic sections are paraphyletic, and three to four growth forms evolved repeatedly. Manihotoides pauciflora is nested within North American Manihot species. Some PEPC and G3pdh clones grouped with clones of other species and not with clones from their own species. CONCLUSIONS: North and South American Manihot species are sister clades. Paraphyly of North American sections suggests that taxonomic revision is warranted. The position of Manihotoides pauciflora confirms that Manihotoides should remain subsumed within Manihot. Most growth forms likely evolved repeatedly in this group. The behavior of PEPC and G3pdhNA clones is probably due to incomplete lineage sorting.

Cytogenetic and anatomic behavior of cytochimeras and total polyploids in cassava.

Cassava periclinal cytochimeras, cultivars, and interspecific hybrid and polyploid types were studied in relation to embryonic, cytogenetic, and anatomical behavior. Their apical shoots, pollen grains, male and female buds, roots, stomata, and flowering period were analyzed. Chimeras exhibited increased size of L1 and L2 cells. Polyploidy led to enlargement of stomata in chimeras whereas L2 gave tetraploid chromosome configurations, tetrad irregularity, decrease of pollen viability, and increase in frequency of polyembryo sacs. The chimeric composition of tetraploids L1 and L2 and diploid L3 expressed a notable epigenetic effect seen in a marked enlargement of edible roots compared to total diploid. One of the chimeric types was accompanied by complete flowering inhibition. Pollen viability and diameter appeared to be reliable markers to determine ploidy levels.

Synthesis of periclinal chimera in cassava.

We provide the first report on the synthesis of a very productive interspecific periclinal chimera of cassava, with large and edible roots. The epidermal tissue of the chimera was formed by the cultivated species Manihot esculenta (E), and the subepidermis and internal tissue were formed by the wild species, Manihot fortalezensis (F). We used cytogenetics and morphological analyses to determine the origins of all tissues. These results may offer potential for the development of new lines for crop improvement based on the use of chimera composed of different combinations of wild species and cultivars.

Assessment of attractiveness of cassava as a roosting plant for the melon fly, Bactrocera cucurbitae, and the Oriental fruit fly, B. dorsalis.

Application of bait spray to crop borders is a standard approach for suppression of melon fly, Bactrocera cucurbitae (Coquillett) (Diptera: Tephritidae) populations and may also be of value for suppression of oriental fruit fly, B. dorsalis (Hendel) populations. Establishment of preferred roosting hosts as crop borders may help to improve suppression of both fruit fly species by providing sites for bait spray applications. In an area-wide B. cucurbitae suppression trial, the question was raised as to whether cassava, Manihot esculenta Crantz (Euphorbiales: Euphorbiaceae), could be used as a B. cucurbitae roosting host. M. esculenta was of interest as a roosting host because, in contrast to many other identified preferred roosting hosts, it would also be a crop potentially increasing the productivity of the crop production system overall. As a short-lived and shrubby perennial, M. esculenta potentially constitutes a crop with more persistent roosting foliage than an annual crop such as corn, Zea mays L. (Cyperales: Poaceae), that has often been planted as a roosting host for B. cucurbitae control. Using protein-baited traps set amidst potted plants placed adjacent to a papaya Carica papaya L. (Violales: Caricaceae) orchard known to have established populations of B. cucurbitae and B. dorsalis, the effectiveness of M. esculenta as a roosting host was assessed by comparing its attractiveness to that of castor bean, Ricinus communis L (Malpighiales: Euphorbiaceae), previously identified as one of the most attractive roosting hosts for B. cucurbitae, and to corn, a crop which has been planted as a roosting host for help in B. cucurbitae control. The results showed that use of M. esculenta as a roosting host is comparable to use of R. communis by both B. cucurbitae and B. dorsalis. These results provide encouragement to incorporate M. esculenta on a farm as a trap crop (i.e. site for bait spray application). This has the advantage of having the trap crop be a crop on its own (as opposed to castor bean) and, among prospective crops that could be used as a trap crop, has foliage more persistent than an annual trap crop such as corn.

Genetic diversity of root anatomy in wild and cultivated Manihot species.

An anatomical study of roots was conducted on two wild Manihot species, namely M. glaziovii and M. fortalezensis, and two cassava varieties, M. esculenta Crantz variety UnB 201 and M. esculenta variety UnB 122, to identify taxonomic differences in primary growth. Anatomical characters of cassava roots have been rarely investigated. Their study may help cassava breeders to identify varieties with economically important characters, such as tolerance to drought. We investigated tap and lateral adventitious roots of two specimens of each clone or species. Free-hand cross-sections of roots were drawn; these had been clarified with 20% sodium hypochlorite solution, stained with 1% safranin-alcian blue ethanolic solution, dehydrated in ethanol series and butyl acetate and mounted in synthetic resin. Anatomical differences among Manihot species and varieties were found in the epidermal and exodermal cell shape and wall thickness, content of cortical parenchyma, and number of xylem poles. Wall thickness of the epidermis and exodermis of tap root were similar in all species, while in the lateral root there were differences in cell shape and wall thickness. Epidermal cells with thick walls were found in the tap root of all species and in lateral roots of cassava varieties. This character is apparently associated with tolerance to drought and disease. The variation in the number of xylem poles of cassava varieties was larger (4-8) than in wild species (4-6), and appears to support the hybrid origin of cassava.

Genetic, embryonic and anatomical study of an interspecific cassava hybrid.

A molecular, anatomical and cytogenetic study of an interspecific hybrid between Manihot esculenta (cassava) and the wild species M. oligantha was carried out. Cytogenetics revealed relatively complete chromosome pairing and high viability of the pollen grains. Ovule structure examined by the clearing method showed polyembryony in 2.7% of the ovules. Doubling of the chromosome number resulted in an increase in polyembryony of up to 18% and a reduction in pollen viability. Multivalent formation was also observed. An anatomical study of stems of diploid and tetraploid hybrids showed a larger number of vascular bundles in the tetraploid type.

Anatomical alterations due to polyploidy in cassava, Manihot esculenta Crantz.

Information on anatomical structure is needed by breeders working on improvement for drought tolerance. For studying the effect of polyploidy on cassava anatomy and its significance to tolerance to drought, we induced a polyploidy type of a selected clone (UnB 530) by applying an aqueous solution of 0.2% colchicine on lateral buds for a period of 12 h. The stem identified as tetraploid was propagated to produce the whole plant. Free-hand cross-sections of the median portion between stem internodes were made. They were clarified using 50% sodium hypochlorite solution, stained with 1% safranin-alcian blue, passed through an ethanol series and butyl acetate and mounted in synthetic resin. The tetraploid type showed more prismatic and druse crystals in the cortical parenchyma, and its pericycle fibers had thicker walls. The secondary xylem of tetraploid types was wider than diploid ones, having thinner walls and less starch.

Wild Manihot species: botanical aspects, geographic distribution and economic value.

A total of 98 Manihot species have been recognized in the genus. All of them are native to the tropics of the New World, particularly Brazil and Mexico. The cultigen, Manihot esculenta Crantz (cassava), grows throughout the lowland tropics. Wild species vary in growth habit from acaulescent or short shrubs to tree-like. Because of their adaptations to different conditions, they are gene reservoirs for tackling many abiotic and biotic stresses such as improving root quality and resistance to diseases. They have been used successfully by the first author for improving protein content, seed-fertility, apomixis, resistance to mealy bug, and tolerance to drought. A table of the most important species from an economic viewpoint is presented.

Wild Manihot species: botanical aspects, geographic distribution and economic value

A total of 98 Manihot species have been recognized in the genus. All of them are native to the tropics of the New World, particularly Brazil and Mexico. The cultigen, Manihot esculenta Crantz (cassava), grows throughout the lowland tropics. Wild species vary in growth habit from acaulescent or short shrubs to tree-like. Because of their adaptations to different conditions, they are gene reservoirs for tackling many abiotic and biotic stresses such as improving root quality and resistance to diseases. They have been used successfully by the first author for improving protein content, seed-fertility, apomixis, resistance to mealy bug, and tolerance to drought. A table of the most important species from an economic viewpoint is presented.

Anatomical alterations due to polyploidy in cassava, Manihot esculenta Crantz

Information on anatomical structure is needed by breeders working on improvement for drought tolerance. For studying the effect of polyploidy on cassava anatomy and its significance to tolerance to drought, we induced a polyploidy type of a selected clone (UnB 530) by applying an aqueous solution of 0.2% colchicine on lateral buds for a period of 12 h. The stem identified as tetraploid was propagated to produce the whole plant. Free-hand cross-sections of the median portion between stem internodes were made. They were clarified using 50% sodium hypochlorite solution, stained with 1% safranin-alcian blue, passed through an ethanol series and butyl acetate and mounted in synthetic resin. The tetraploid type showed more prismatic and druse crystals in the cortical parenchyma, and its pericycle fibers had thicker walls. The secondary xylem of tetraploid types was wider than diploid ones, having thinner walls and less starch.

Ocorrência de isômeros do beta-caroteno em raízes de cinco cultivares de mandioca (Manihot esculenta Crants) do Estado de São Paulo / Beta-carotene isomers in roots of five manioc (Manihot esculenta Crantz) cultivars of São Paulo State

Raízes de cinco cultivares de mandioca (Manihot esculenta Crantz), provenientes do Instituto Agronômico de Campinas, a saber: Branca de Santa Catarina (SRT-59), Pioneira (SRT-13l0), Ouro do Vale (SRT-797), IAC-576-70 e IAC 289-70. As análises visaram a identificação dos principais carotenóides presentes nas raízes dos cultivares de mandioca produzidos no Estado de são Paulo, bem como a determinação da atividade pró-vitamínica A dos mesmos. Pelo cozimento das raízes ou a transformação em farinha, foi possível avaliar as perdas do teor vitamínico A de correntes do processamento. As análises foram realizadas no período de julho/86 a janeiro/87, sendo, desta maneira, possível avaliar as prováveis perdas decorrentes do armazenamento. Os resultados das análises permitiram concluir que os principais carotenóides presentes nas raízes dos cultivares de mandioca estudados foram o neo-ß-caroteno B, o ß-caroteno todo¬-trans e o neo-ß-caroteno U. |Os teores de vitamina A nas mandiocas estudadas, expressos em equivalentes de retinol/l00g, variaram de 2,8 a 13,9 para as amostras cruas e de 0,4 a 10,7 para as amostras processadas, sendo que o cozimento promoveu uma diminuição da atividade pró-vitamínica A, que variou de 20 a 55%. Dos cultivares estudados o IAC 289-70 foi o que apresentou menor porcentagem de perda de atividade pró-vitamínica A com o cozimento. Quanto ao armazenamento, foi possível observar que a mandioca crua com casca armazenada por 6 meses em freezer a –20°C não apresentou perdas na sua atividade pró-vitamínica A enquanto que o armazenamento das farinhas à temperatura ambiente, ao abrigo da luz, pelo mesmo período de tempo, apresentou total degradação de seus carotenóides

Roots of five following cultivars of cassava (Manihot esculenta Crantz) from "Instituto Agronômico de Campinas" were analyzed: "Branca de Santa Catarina" (SRT-59), "Pioneira" (SRT-1310), "Ouro do Vale" (SRT-797), "IAC 576-70" and "IAC 289-70". The analyses were made for the identification of the main carotenoids present in roots of the cassava cultivars produced in São Paulo State, as well as for determination of their pro-vitamin A activity. The cooking of the roots or their transformation as flour made possible to evaluate the loss of the pro-vitamin A value which occurs with processing. The analyses were made from July/1986 to January/1987. This permitted to evaluate the possibility of losses during storage. The results of the analyses allowed to conclude that the main carotenoids present in roots of the studied cassava cultivars were neo-ß-carotene B, all-trans-ß-carotene and neo-ß-carotene U. The vitarnin A values in these cassava expressed as retinol equivalents/lOOg varied from 2,8 to 13,9 for the raw samples; and with cooking, a diminution of the pro-vitarnin A activity occurred, which varied from 20 to 55%.Of all the studied cultivars, the "IAC 289-70" was the one that showed the greatest percentages of loss of pro-vitamin A activity with the cooking. The raw cassavas stored at –20°C for 6 months didn't show losses of their pro-vitamin A value, while the storage of the flour, at room temperature with no light, for the same period of time, showed total degradation of their carotenoids