Flower-inducing technology facilitates speed breeding in cassava.

Cassava is a tropical crop that provides daily carbohydrates to more than 800 million people. New cassava cultivars with improved yield, disease resistance, and food quality are critical to end hunger and reduce poverty in the tropics. However, the progress of new cultivar development has been dragged down by difficulties obtaining flowers from desired parental plants to enable designed crosses. Inducing early flowering and increasing seed production are crucial to improving the efficiency of developing farmer-preferred cultivars. In the present study, we used breeding progenitors to evaluate the effectiveness of flower-inducing technology, including photoperiod extension, pruning, and plant growth regulators. Photoperiod extension significantly reduced the time to flowering in all 150 breeding progenitors, especially late-flowering progenitors which were reduced from 6-7 months to 3-4 months. Seed production was increased by using the combination of pruning and plant growth regulators. Combining photoperiod extension with pruning and the PGR 6-benzyladenine (synthetic cytokinin) produced significantly more fruits and seeds than only photoperiod extension and pruning. Another growth regulator, silver thiosulfate, commonly used to block the action of ethylene, did not show a significant effect on fruit or seed production when combined with pruning. The present study validated a protocol for flower induction in cassava breeding programs and discussed factors to consider in implementing the technology. By inducing early flowering and increasing seed production, the protocol helped move one step further for speed breeding in cassava.

Native and fermented waxy cassava starch as a novel gluten-free and clean label ingredient for baking and expanded product development.

Amylose-free and wild-type cassava starches were fermented for up to 30 days and oven- or sun-dried. The specific volume (ν) after baking was measured in native and fermented starches. The average ν (across treatments) for waxy starch was 3.5 times higher than that in wild-type starches (17.6 vs. 4.8 cm3 g-1). The best wild-type starch (obtained after fermentation and sun-drying) had considerably poorer breadmaking potential than native waxy cassava (8.4 vs. 16.4 cm3 g-1, respectively). The best results were generally obtained through the synergistic combination of fermentation (for about 10-14 days) and sun-drying. Fermentation reduced viscosities and the weight average molar mass led to denser macromolecules and increased branching degree, which are linked to a high loaf volume. The absence of amylose, however, was shown to be a main determinant as well. Native waxy starch (neutral in taste, gluten-free, and considerably less expensive than the current alternatives to cassava) could become a new ingredient for the formulation of clean label-baked or fried expanded products.

Correlation of cooking time with water absorption and changes in relative density during boiling of cassava roots.

Consumers prefer cassava roots that cook quickly during boiling. Current methods to evaluate cooking time (CT) are slow and labour-intensive. This article describes improved protocols for assessing CT in roots. We evaluated CT in 36 genotypes monthly at 8-11 months after planting. CT showed differences for plant age at harvest and among genotypes. During boiling, roots absorbed water (WAB) and thus reduced their relative density (DEN). We classified three groups of genotypes with increasing CT (≤25 min, 25-40 min and >40 min), associated with decreasing WAB, respectively, 15.3 ± 3.1, 10.7 ± 1.7 and 4.9 ± 3.8% of initial root weight. A similar trend was observed for changes in DEN (46.3 ± 9.8, 54.5 ± 11.1 and 75.9 ± 6.9% of initial DEN, respectively). The highest correlations between WAB and DEN with CT (r 2 > 0.6) were found at 30-min boiling. These alternative protocols facilitate screening large numbers of cassava genotypes for CT.

Effect of Pruning Young Branches on Fruit and Seed Set in Cassava.

Flowering in cassava is closely linked with branching. Early-flowering genotypes branch low and abundantly. Although farmers prefer late flowering genotypes because of their erect plant architecture, their usefulness as progenitors in breeding is limited by their low seed production. In general, the first inflorescence aborts in cassava. Preventing this abortion would result in early production of seeds and make cassava breeding more efficient. The objective of this study was to assess if pruning young branches prevents the abortion of first inflorescences and promotes early fruit and seed set. Four genotypes with early, late, very late, and no flowering habits were grown under an extended photoperiod (EP) or normal dark night conditions (DN). Additional treatments included pruning young branches at the first or second flowering event and spraying (or not) benzyladenine (BA) after pruning. One genotype failed to flower and was not considered further. For the remaining genotypes, EP proved crucial to induce an earlier flowering, which is a pre-requisite for pruning. Total production of seeds in EP plots was 2,971 versus 150 in DN plots. For plants grown under EP, the average number of seeds per plant without pruning was 3.88, whereas those pruned produced 17.60 seeds per plant. Pruning at the first branching event led to higher number of seeds per plant (26.25) than pruning at the second flowering event (8.95). In general, applying BA was beneficial (38.52 and 13.98 seeds/plant with or without spraying it, respectively). The best combination of treatments was different for each genotype. Pruning young branches and applying BA in the first flowering event not only prevented the abortion of inflorescences but also induced the feminization of male flowers into hermaphrodite or female-only flowers. The procedures suggested from this study (combining EP, pruning young branches, and spraying BA), allowed the production of a high number of seeds from erect cassava genotypes in a short period. The implementation of these procedures will improve the breeding efficiency in cassava.

Discovery of an amylose-free starch mutant in cassava (Manihot esculenta Crantz).

One of the objectives of the cassava-breeding project at CIAT is the identification of clones with special root quality characteristics. A large number of self-pollinations have been made in search of useful recessive traits. During 2006 harvests an S1 plant produced roots that stained brownish-red when treated with an iodine solution, suggesting that it had lower-than-normal levels of amylose in its starch. Colorimetric and DSC measurements indicated low levels (3.4%) and an absence of amylose in the starch, respectively. SDS-PAGE demonstrated the absence of GBSS enzyme in the starch from these roots. Pasting behavior was analyzed with a rapid visco-analyzer and resulted in larger values for peak viscosity, gel breakdown, and setback in the mutant compared with normal cassava starch. Solubility was considerably reduced, while the swelling index and volume fraction of the dispersed phase were higher in the mutant. No change in starch granule size or shape was observed. This is the first report of a natural mutation in cassava that drastically reduces amylose content in root starch.

Biological Implications in Cassava for the Production of Amylose-Free Starch: Impact on Root Yield and Related Traits.

Cassava (Manihot esculenta, Crantz) is an important food security crop, but it is becoming an important raw material for different industrial applications. Cassava is the second most important source of starch worldwide. Novel starch properties are of interest to the starch industry, and one them is the recently identified amylose-free (waxy) cassava starch. Waxy mutants have been found in different crops and have been often associated with a yield penalty. There are ongoing efforts to develop commercial cassava varieties with amylose-free starch. However, little information is available regarding the biological and agronomic implications of starch mutations in cassava, nor in other root and tuber crops. In this study, siblings from eight full-sib families, segregating for the waxy trait, were used to determine if the mutation has implications for yield, dry matter content (DMC) and harvest index in cassava. A total of 87 waxy and 87 wild-type starch genotypes from the eight families were used in the study. The only significant effect of starch type was on DMC (p < 0.01), with waxy clones having a 0.8% lower content than their wild type counterparts. There was no effect of starch type on fresh root yield (FRY), adjusted FRY and harvest index. It is not clear if lower DMC is a pleiotropic effect of the waxy starch mutation or else the result of linked genes introgressed along with the mutation. It is expected that commercial waxy cassava varieties will have competitive FRYs but special efforts will be required to attain adequate DMCs. This study contributes to the limited knowledge available of the impact of starch mutations on the agronomic performance of root and tuber crops.

Cassava Breeding II: Phenotypic Correlations through the Different Stages of Selection.

Breeding cassava relies on a phenotypic recurrent selection that takes advantage of the vegetative propagation of this crop. Successive stages of selection (single row trial-SRT; preliminary yield trial-PYT; advanced yield trial-AYT; and uniform yield trials UYT), gradually reduce the number of genotypes as the plot size, number of replications and locations increase. An important feature of this scheme is that, because of the clonal, reproduction of cassava, the same identical genotypes are evaluated throughout these four successive stages of selection. For this study data, from 14 years (more than 30,000 data points) of evaluation in a sub-humid tropical environment was consolidated for a meta-analysis. Correlation coefficients for fresh root yield (FRY), dry matter content (DMC), harvest index (HIN), and plant type score (PTS) along the different stages of selection were estimated. DMC and PTS measured in different trials showed the highest correlation coefficients, indicating a relatively good repeatability. HIN had an intermediate repeatability, whereas FRY had the lowest value. The association between HIN and FRY was lower than expected, suggesting that HIN in early stages was not reliable as indirect selection for FRY in later stages. There was a consistent decrease in the average performance of clones grown in PYTs compared with the earlier evaluation of the same genotypes at SRTs. A feasible explanation for this trend is the impact of the environment on the physiological and nutritional status of the planting material and/or epigenetic effects. The usefulness of HIN is questioned. Measuring this variable takes considerable efforts at harvest time. DMC and FRY showed a weak positive association in SRT (r = 0.21) but a clearly negative one at UYT (r = -0.42). The change in the relationship between these variables is the result of selection. In later stages of selection, the plant is forced to maximize productivity on a dry weight basis either by maximizing FRY or DMC, but not both. Alternatively, the plant may achieve high dry root yield by simultaneously attaining "acceptable" (but not maximum) levels of FRY and DMC.

Cassava Breeding I: The Value of Breeding Value.

Breeding cassava relies on several selection stages (single row trial-SRT; preliminary; advanced; and uniform yield trials-UYT). This study uses data from 14 years of evaluations. From more than 20,000 genotypes initially evaluated only 114 reached the last stage. The objective was to assess how the data at SRT could be used to predict the probabilities of genotypes reaching the UYT. Phenotypic data from each genotype at SRT was integrated into the selection index (SIN) used by the cassava breeding program. Average SIN from all the progenies derived from each progenitor was then obtained. Average SIN is an approximation of the breeding value of each progenitor. Data clearly suggested that some genotypes were better progenitors than others (e.g., high number of their progenies reaching the UYT), suggesting important variation in breeding values of progenitors. However, regression of average SIN of each parental genotype on the number of their respective progenies reaching UYT resulted in a negligible coefficient of determination (r (2) = 0.05). Breeding value (e.g., average SIN) at SRT was not efficient predicting which genotypes were more likely to reach the UYT stage. Number of families and progenies derived from a given progenitor were more efficient predicting the probabilities of the progeny from a given parent reaching the UYT stage. Large within-family genetic variation tends to mask the true breeding value of each progenitor. The use of partially inbred progenitors (e.g., S1 or S2 genotypes) would reduce the within-family genetic variation thus making the assessment of breeding value more accurate. Moreover, partial inbreeding of progenitors can improve the breeding value of the original (S0) parental material and sharply accelerate genetic gains. For instance, homozygous S1 genotypes for the dominant resistance to cassava mosaic disease (CMD) could be generated and selected. All gametes from these selected S1 genotypes would carry the desirable allele and 100% of their progenies would be resistant. Only half the gametes produced by the heterozygous S0 progenitor would carry the allele of interest. For other characteristics, progenies from the S1 genotypes should be, at worst, similar to those generated by the S0 progenitors.

Induction and identification of a small-granule, high-amylose mutant in cassava (Manihot esculenta Crantz).

Only two mutations have been described in the literature, so far, regarding starch and root quality traits in cassava. This article reports on an induced mutation in this crop, first identified in 2006. Botanical seed from five different cassava families were irradiated with gamma rays. Seed was germinated, transplanted to the field (M1 plants), and self-pollinated to produce the M2 generation. Abnormal types regarding starch granule morphology were identified during the single plant evaluation of M2 genotypes. To confirm these characteristics, selected genotypes were cloned and a second evaluation, based on cloned plants obtained from vegetative multiplication, was completed in September 2007. Two M2 genotypes presented small starch granules, but only one could be fully characterized, presenting a granule size of 5.80 +/- 0.33 microm compared with three commercial clones with granule sizes ranging from 13.97 +/- 0.12 to 18.73 +/- 0.10 microm and higher-than-normal amylose content (up to 30.1% in cloned plants harvested in 2007, as compared with the typical values for "normal" cassava starch of around 19.8%). The gels produced by the starch of these plants did not show any viscosity when analyzed with the rapid viscoanalyzers (5% suspension), and the gels had low clarity. Low viscosity could be observed at higher concentrations (8 or 10% suspensions). Preliminary results suggest that the mutation may be due to a lesion in a gene encoding one of the isoforms of isoamylase (probably isa1 or isa2).