Apomixis in different ploidy levels of cassava.

Two polyploid hybrids between cassava (Manihot esculenta) cultivar 307-2 and its wild relatives M. glaziovii and M. anomala, were studied to examine the relationship between ploidy level and the production of seeds without fertilization. A clearing method was applied to assess ovule sizes as an indication of multiembryony. The diploid cultivar 307-2 had regular 18 bivalents at meiotic metaphase 1 while the polyploid types showed chromosome configurations varying from 3 to 4 quadrivalents and 28 to 30 bivalents. A total of 14% of studied ovules of the polyploid hybrid involving M. glaziovii were multiebryonic, while the percentage of multiembryony was as low as 2% in the polyploid hybrid M. anomala×M. esculenta. Diploid hybrid types did not show any multi embryony. Adventitious embryos were found and documented for the first time in polyploid hybrids M. esculenta×M. glaziovii. The association of multiple embryo formation with ovary size and pollination showed that apomictic embryos form independently from fertilization. Simple iodized carmine stain for measuring pollen viability proved as efficient as the sophisticated Alexander method.

Alterations of reproduction system in a polyploidized cassava interspecific hybrid.

The objective of this research was to examine how much polyploidy may affect seed and root formation in cassava interspecific hybrids Manihot esculenta Crantz xM. oligantha Pax. A polyploid type was induced by colchicine treatment to lateral buds followed by propagating vegetatively arising stems. Cytogenetic and anatomical analyses were made on both polyploid and diploid types. The polyploid type showed extensive chromosome pairing and pollen viability. Multiembryonic ovule frequency increased in polyploid plants. Stalks became woody and propagation through roots difficult, the edible roots increased, however, in size.

Improving carotenoids and amino-acids in cassava.

More than 800 million people in tropics and sub tropics use cassava as food. However, its roots are poor in protein content (0.7-2%). Amino acids such as lysine and methionine are also low, and some research reports indicate the absence of methionine in cassava edible roots. By inter-specific hybridization it was possible to increase true protein in cassava roots measured by amino acid contents. The amino acid profiles of a common cassava cultivar and an inter-specific hybrid, namely ICB 300, were determined using the computerized amino acid analyzer Hitachi L-8500. The inter-specific hybrid has 10-fold lysine and 3-fold methionine than common cassava cultivar: lysine content was 0.010 g per 100 g in the common cassava cultivar while it reached 0.098 in the inter-specific hybrid. Methionine in the common cassava cultivar was 0.014 g per 100 g whereas it reached 0.041 g per 100 g in the inter-specific hybrid. Total amino acid content in the common cassava cultivar was 0.254 g per 100 g viz. a viz. 1.664 g per 100 g in the inter-specific hybrid. The genetic variability of the profile and quantity of amino acids indicate the feasibility of selecting inter-specific hybrids that are rich in both crude protein and amino acids. Carotenoid content could be improved in cassava edible roots by selecting cultivars rich in carotenoids. In Brazil, the center of cassava origin, cassava landraces have acquired through their domestication a large diversity in relation to many economic traits such as high content of carotenoids and excellent palatability among other characters. One of these clones, which has been grown by indigenous farmers in Brazil and available at the University of Brasília genebank, showed a high level of lycopene content (5 mg/kg viz. a viz. zero in common cultivars, and 12-20 mg/kg in tomato-a lycopene-rich vegetable). The cassava landrace UnB 400 had a high content of beta-carotene (up to 4 mg/kg). This article also discusses relevant patents to the main subject of this research.

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.

A further study of microsatellite on apomixis in cassava.

Cassava is the main staple crop in the tropics. It is vegetatively propagated by stem cuttings that maintain superior genotypes, but favors disease accumulation. In this report we present the results of the screening of the progeny and the second generation of the clone UnB 307 for apomixis using microsatellites. A total of 29 plants were screened, representing the maternal plant, its first and second generations, that were left to open pollination. About 20% of the offspring were rated as genetically identical plants. This result confirms the facultative apomictic nature of cassava, with high environment effect.

Amino acid profile in cassava and its interspecific hybrid.

Cassava roots have a low-protein content (0.7-2%). Amino acids such as lysine and methionine are also low, and some research reports have indicated the absence of methionine. The amino acid profiles of a common cassava cultivar and an interspecific hybrid, namely ICB 300, were determined using the computerized amino acid analyzer Hitachi L-8500. The interspecific hybrid has 10 times more lysine and 3 times more methionine than the common cassava cultivar: lysine content was 0.010 g per 100 g in the common cassava cultivar while it reached 0.098 in the interspecific hybrid. Methionine in the common cassava cultivar was 0.014 g per 100 g whereas it reached 0.041 g per 100 g in the interspecific hybrid. Total amino acid content in the common cassava cultivar was 0.254 g per 100 g viz. a viz. 1.664 g per 100 g in the interspecific hybrid. The genetic variability of the profile and quantity of amino acids indicate the feasibility of selecting interspecific hybrids that are rich in both crude protein and amino acids. This is the first report of high true protein in cassava root.

Cassava genetic resources and their utilization for breeding of the crop.

Wild cassava relatives are perennials and vary in growth pattern from nearly acaulescent subshrubs to small trees. They have been used as a source of useful characters such as high protein content, apomixis, resistance to mealybug and mosaic disease, and tolerance to drought. Indigenous clones are a potential source of beta-carotene and lycopene. Apomixis genes have been transferred to the crop successfully through interspeci fi c hybridization, and apomictic clones arising from these hybrids are now being grown at the Universidade de Brasília. Interspeci fi c hybrids produced earlier were polyploidized and had their fertility restored. Different useful types of chimera were also produced.

Cassava in South America, Brazil's contribution and the lesson to be learned from India.

South America is responsible for about half of the cassava world production. In the 1970's productivity of the crop on the continent was about 15 ton/ha, and dropped continuously until reaching 12 ton/ha in 2004. India's productivity of cassava increased from 10 ton/ha in the 1970's to 28 ton/ha in 2004. Brazil contributed significantly to improving cassava crops through the Instituto Agronômico de Campinas in the 1960's and 1970's. The Universidade de Brasília released high-protein content hybrids, apomictic clones and explored the potential of indigenous landraces.

Chromosome doubling induces apomixis in a cassava x Manihot anomala hybrid.

A new species was synthesized artificially by chromosome doubling of an interspecific hybrid between cassava and Manihot anomala. The ensuing polyploid type exhibits an apomictic nature and maintains its morphological characteristics in the progeny. It showed a 29% frequency of multiembryonic sacs in its examined ovules whereas the multiembryonic sacs were absent in the diploid type.

Microsatellite markers confirm high apomixis level in cassava bred clones.

Apomixis genes have successfully been transferred to cassava by hybridization with the wild species, Manihot glaziovii. The interspecific hybrid of cassava and M. glaziovii was left for open pollination during the subsequent three generations. Seven sibs and their maternal progenitor of the fourth generation were genotyped using five microsatellite loci previously developed for cassava. All sibs were identical with each other and with their maternal progenitor. Sibs from M. glaziovii itself proved to be identical when examined by the same microsatellite loci. These results lead to the conclusion that apomixis do occur in wild cassava relatives and apparently has played an important role in Manihot speciation.

Breeding cassava for apomixis.

Apomixis genes have been successfully transferred to cassava (Manihot esculenta) by hybridizing it with the wild species, M. glaziovii. The interspecific hybrid of cassava and M. glaziovii was exposed to open pollination during three subsequent generations. Seven sibs and the maternal progenitor of the fourth generation were genotyped using five microsatellite loci previously developed for cassava. All sibs were identical with each other and with their maternal progenitor. Sibs from M. glaziovii proved to be identical when examined by the same microsatellite loci. This evidence leads to the conclusion that apomixis does occur in wild-cassava relatives and apparently has played an important role in Manihot speciation. This is the first report of nearly 100% apomixis.

Cassava, Manihot esculenta Crantz genetic resources: VI. Anatomy of a diversity center.

Central Brasil is one of the four centers of diversity of Manihot species. Of 26 wild Manihot species reported to occur in this region, 20 were collected from two limited areas, each less than 100 km in diameter. These two areas are Goiás Velho and Corumbá de Goiás. Considering the Harlan concept of geographic patterns of variation of cultivated crops, it is assumed that the two areas are microcenters of cassava diversity. Topographic, soil and ethnological data of these species were collected and compared.

Gene flow between cassava, Manihot esculenta Crantz, and wild relatives.

Controlled and natural hybridization between cassava and wild relatives does occur. Barriers within the genus appear to be weak due to recent evolution of the group. All Manihot species examined cytogenetically have a chromosome number of 2n = 36. However, they behave meiotically as diploids. The weak interspecific barriers have led to an extremely heterozygous gene pool that may begin a sequence of hybridization followed by speciation. Introgression from cassava into a number of wild species (M. neusana, M. alutacea, M. reptans and M. anomala) has been detected by both morphological marker genes and molecular techniques. Winged fruit, setaceous bracteoles, and wide leaf sinus were dominant genes that came from cassava and appeared in the hybrids. The characteristic protein bands of cassava were recognized in the hybrid seed protein electrophoresis.

Cassava, Manihot esculenta Crantz, genetic resources: origin of the crop, its evolution and relationships with wild relatives.

About 98 species of Manihot are known. All of them are native to the New World and are concentrated in four regions in Brazil and Central America. All the Manihot species so far examined have 2n = 36 chromosomes. Interspecific hybrids between cassava and its wild relatives show relatively normal meiosis, and further generations can be obtained. Electrophoresis shows affinity among wild species of different sections, and between some of them and cassava. Both polyploidy and apomixis may have contributed to speciation in this genus. Polyploidy produced genetic variability, while apomixis is responsible for perpetuating new hybrid types adapted to different environments. Cassava may have originated by hybridization between two wild Manihot species, followed by vegetative reproduction of the hybrid.

Apomixis and cassava.

Apomixis means seed formation without fertilization. In cassava (Manihot esculenta) it is an alternative to reproduction by cuttings, which normally transmits pathogens and leads to an accumulation of viral and bacterial diseases. Apomixis also assures preservation of heterosis and avoids genetic segregation. It occurs in wild relatives of cassava and has been transferred successfully from Manihot glaziovii and M. neusana. It is facultative, and occurs at a low frequency, ranging from 1-2%, and apparently is genetically different from apomixis in other crops. With selection, the frequency can reach 13%. Apomixis in cassava is frequently associated with aneuploidy but it does occur in some diploid types. It is due to the formation of aposporic sacs, which can easily be detected by clearing tissue preparations. Apomixis appears to have played an important role in speciation during the evolution of Manihot, since it leads to the maintenance and perpetuation of sterile interspecific hybridization. The use of apomixis in cassava breeding could lead to a boom in line improvement and commercial production. In addition to preserving superior genotypes, avoiding contamination of new plants, it would enable international programs to export their germplasm to destination countries. This would allow the use of superior genotypes even if apomixis occurs at a low frequency. A scheme to maximize benefits is to use diploid apomictic clones as maternal parents, which can be crossed with pollinators of polyploid interspecific hybrids, followed by selection among the progeny of new apomictic types that combine the heteroses of both interspecific hybridization and polyploidy. In addition, they acquire favored genes that have been transferred from the wild to the commercial crop.