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).     

Novel ssIIa Alleles Produce Specific Seed Amylose Levels in Hexaploid Wheat

Novel starch synthase II (ssIIa ) alleles were created in a soft wheat (Triticum aestivum L.) via ethyl methanesulfonate mutagenesis and combined by crossing to create four unique ssIIa triple mutant haplotypes. A range of starch amylose content was obtained from the ssIIa triple mutant lines (35.6–46.8%), with moderate increases in amylose content found in lines carrying two ssIIa null alleles and one partially functional ssIIa allele. Seeds from all ssIIa triple mutants had significantly higher amylose and protein contents but also lower starch content, kernel weight, and flour swelling power compared with their wild‐type sister lines. Seed starch amylose content was negatively correlated with individual kernel weight (r = –0.54), starch content (r = –0.85), and flour swelling power (r = –0.91) but positively correlated with grain protein (r = 0.78), demonstrating that unique ssIIa triple mutant null combinations confer defined levels of seed starch amylose, protein content, and kernel size. The ssIIa mutant lines also had a hard grain texture (≈86 single kernel characterization system units), whereas all parental material and wild‐type sister lines had soft grain texture (≈35). This change in grain texture was independent of Ha locus genotype, because all lines carried a functional (soft type) Ha locus. The ssIIa alleles and allelic combinations reported here demonstrate the ability to create defined levels of seed starch, amylose, and protein.     

The effect of vesicular-arbuscular mycorrhizal inoculation on nutrient uptake and yield of alley-cropped cassava in a degraded Alfisol of southwestern Nigeria

Leaf and root (tuber) nutrient uptake patterns of cassava (Manihot esculenta Crantz) alley-cropped with gliricidia (Gliricidia sepium), leucaena (Leucaena leucocephala), and senna [(Senna (syn. Cassia) siamea] as influenced by vesicular-arbuscular mycorrhizal (VAM) inoculation in a degraded Alfisol were investigated in consecutive years. The cassava plants were mulched with fresh prunings of each hedgerow tree species at 2-month intervals in the second and third years of alley cropping. While VAM inoculation significantly influenced the root uptake of nutrients, the leaf uptake was not affected except for the uptake of P. In most cases, there was no difference in the nutrient concentration between inoculated and uninoculated plants, either in the leaf or in the root, indicating that the productivity of cassava was regulated by the amount of nutrients the roots could absorb. In spite of similar total soil N in all inoculated and uninoculated alley-cropped cassava plots and similar exchange-able soil K contents in inoculated and uninoculated alley-cropped cassava plots with leucaena and senna, greater uptake of N, P, and K and greater concentrations of K were observed in roots of inoculated alley-cropped cassava with gliricidia and leucaena than with senna. These results indicated that greater mineralization and availability of nutrients to cassava roots from prunings of nodulating gliricidia and leucaena than from non-nodulating senna may be important, particularly with efficient VAM inoculation, in these alley-cropping systems. Also, for similar nutrients in the inoculated and uninoculated cassava soils alley-cropped with each hedgerow species, VAM inoculation significantly enhanced cassava root dry weights, indicating that an effective VAM fungus can be an agent of greater nutrient uptake in a competitive environment.     

Structural characterization of Peruvian carrot (Arracacia xanthorrhiza) starch and the effect of annealing on its semicrystalline structure

Structural characteristics of native and annealed Peruvian carrot (Arracacia xanthorrhiza) starches were determined and compared to those of cassava and potato starches. Peruvian carrot starch presented round and irregular shaped granules, low amylose content and B-type X-ray pattern. Amylopectin of this starch contained a large proportion of long (DP > 37) and short (DP 6-12) branched chains. These last ones may contribute to its low gelatinization temperature. After annealing, the gelatinization temperatures of all starches increased, but the ΔH and the crystallinity increased only in Peruvian carrot and potato starches. The annealing process promoted a higher exposure of Peruvian carrot amylose molecules, which were more quickly attacked by enzymes, whereas amylopectin molecules became more resistant to hydrolysis. Peruvian carrot starch had structural characteristics that differed from those of cassava and potato starches. Annealing affected the semicrystalline structure of this starch, enhancing its crystallinity, mainly due to a better interaction between amylopectin chains.     

Increase in Apparent Amylose Content and Change in Starch Pasting Properties at Cool Growth Temperatures in Mutant Wheat

Some mutant wheat lines with low‐amylose content were grown in a field and greenhouse (15 or 20°C) to compare apparent amylose content and starch pasting properties. The apparent amylose content of flour and starch increased and starch pasting parameters as measured by a Rapid Visco Analyser (RVA) changed in the greenhouse (at cool temperatures) during seed maturation. Densitometric analysis of the protein band separated by electrophoresis suggested that the increase in amylose content by cool temperature was related to the amount of Wx‐D1 protein. This data suggests that the Wx‐D1 gene was responsible for these changes. In wheat starch from Tanikei A6099 and Tanikei A6598 at 15°C, the value of final viscosity and total setback was higher than that from the field. In wheat starch from Tanikei A6599‐4 (waxy mutant with stable hot paste viscosity), the peak viscosity temperature was higher and time maintained >80% of the peak was shorter at 15°C than that from the field. Genetic analysis using doubled‐haploid (DH) lines from a combination of Tanikei A6599‐4 and Kanto 118 (low‐amylose line) showed that apparent amylose content increased and the starch pasting curve and properties changed in waxy progenies similar to Tanikei A6599‐4.     

Yield and nutritional requirements of cassava in response to potassium fertilizer in the second cycle

Potassium (K) is one of the most absorbed nutrients by cassava because it acts on the synthesis and starch accumulation in the storage roots. Here, we show that K application at the beginning of the second vegetative cycle of cassava submitted to shoot pruning increased the yield of roots and starch, and the nutrient demand of plants. Application of 45–89 kg ha^?1 dipotassium oxide (K₂O) in the second cycle increased the yield of storage roots and starch from 36–49% and K applied at this time had a greater effect on the synthesis and allocation of starch in the storage roots. K supply increased the accumulation of N and S by 2.0- to 3.0-fold and the accumulation of other nutrients by 1.4- to 1.7-fold. The removal of phosphorus (P), manganese (Mn), and zinc (Zn) by storage roots was not affected by K application, whereas the removal of other nutrients increased by 1.3- to 4.3-fold.     

木薯SBEI基因片段克隆及块根特异性反义表达载体的构建

为了抑制木薯支链淀粉的合成,提高直链淀粉含量,改变木薯淀粉结构,培育工业用燃料酒精型木薯品种,本研究利用RT-PCR方法,用木薯块根cDNA克隆获得支链淀粉合成的关键酶基因SBEI的部分片段,对其进行序列分析表明与GenBank序列高度同源,同源性为99.22％;并以pBI121为基础,构建了以块根特异性表达启动子Sporamin驱动的SBEI基因反义结构的植物表达载体。     

Amylose-Lipid Complex Formation in Acetylated Pea Starch-Lipid Systems

Starch-lipid interactions involving native and acetylated pea starch were studied by differential scanning calorimetry (DSC) and measurements of iodine affinity. Lipids including lauric acid, monopalmitin, and butterfat were added to aqueous starch dispersions after the starch was gelatinized at 85°C. DSC thermal curves of gelatinized modified pea starch systems containing fatty acid or monoglyceride did not show DSC transitions indicative of amylose complexes with external lipids, whereas a DSC endotherm of amylose-lipid complexes was observed for the corresponding native starch-lipid systems. However, iodine binding studies revealed that acetylated pea starch amylose complexed with added fatty acid or monoglyceride in the modified pea starch-lipid composites. The failure of DSC detection of the complexes in these systems was attributed to the absence of crystalline structures of acetylated pea starch amylose complexes. Furthermore, acetylation of starch decreased the complexing ability of the pea starch amylose as revealed by a reduction in iodine affinity. Both DSC and iodine affinity studies showed that neither native nor modified pea starch interacted to a significant extent with butterfat that consisted mainly of triglycerides.     

Structural Properties of Starch in Bread and Bread Model Systems: Influence of an Antistaling α‐Amylase

The influence of an antistaling α‐amylase on bread crumb and on wheat starch gels was investigated taking into account different levels of structural hierarchy. Bread was prepared by a conventional baking procedure. Starch gels were produced by heating a concentrated starch dispersion in closed molds. Bread and starch gels were characterized by compression tests, light microscopy (LM), differential scanning calorimetry, and X‐ray measurements. The α‐amylase enhanced the initial firmness of starch gels and reduced the firming rate of bread and starch gels on aging. LM revealed that amylose and amylopectin phase‐separated within the starch granules and that freshly baked control bread and starch gels showed weak birefringence which became more intense during aging. Amylase‐containing bread and starch gels exhibited strong birefringence in the amylose rich region of the granules directly after baking which did not significantly increase during aging. The enzyme hindered the retrogradation of amylopectin as detected by differential scanning calorimetry, whereas X‐ray diffraction indicated that the enzyme induced low levels of starch crystallinity which did not change during aging. It is hypothesized that the antistaling effect of the amylase is based on the capacity to partially degrade amylopectin and, by this, to hinder its recrystallization. On the other hand, the enzyme slightly degrades amylose by an endo‐mechanism which, in turn, promotes the rapid formation of a partly crystalline amylose network in fresh bread and hinders amylose rearrangements during aging.     

EFFECT OF SWINE MANURE EXTRACT ON LEAF NITROGEN CONCENTRATION,CHLOROPHYLL CONTENT,TOTAL POTASSIUM IN PLANT PARTS AND STARCH CONTENT IN FRESH TUBER YIELD OF CASSAVA

The effects of swine manure extract (SME) as foliar fertilizer (FSME), soil fertilizer (SSME), and both soil and foliar fertilizer (FNSSME) on leaf nitrogen (N) concentration, chlorophyll content, total potassium (K) in plant parts and starch content in the fresh roots, compared to a conventional, chemical fertilizer (CF) were studied in cassava cultivar ‘Hauybong 60’. The results showed that plants on FNSSME had the highest chlorophyll contents and SPAD values at four months after planting (MAP). Cassava plants treated with SSME and FNSSME had highest starch content and was significantly higher than in plants treated with CF and a FSME. There was a strong, positive relationship between the leaf nitrogen concentration and chlorophyll content in cassava at 6 MAP. The results of the study indicated that an application of FNSSME to cassava plants could provide a higher chlorophyll content and higher tuber quality of the plants than those applied with chemical fertilizer.     

IBA和MET单独与配合使用对木薯产量和淀粉含量的影响

在50mg／L吲哚丁酸(IBA)浸种的基础上，于块根膨大期配合叶面喷施500mg／L和1000mg／L多效唑(MET)，结果表明，可使木薯产量比对照分别增加18．8％和20．5%．块根淀粉含量则相应提高6．8%和9．0％，均达0．05显著水平；而单独用50mg／L IBA浸种或只在块根膨大期叶面喷施500mg／L和1000mg／L MET，木薯产量和淀粉含量也有不同程度的提高，但效果均不及IBA和MET的配合使用。     

Starch Molecular Mass and Size by Size-Exclusion Chromatography in DMSO-LiBr Coupled with Multiple Angle Laser Light Scattering

The weight average molar mass (M_w) and root mean square radii of starches from waxy maize (Amioca), waxy rice flour, cassava, Hylon V, Hylon VII, and potato amylose were determined by size-exclusion chromatography (SEC) and multiple-angle laser light scattering (MALLS). Dimethylsulfoxide (DMSO) containing 50 mM LiBr was used to dissolve the starches and also served as the mobile phase. SEC with large particle size polystyrene divinylbenzene packing materials and MALLS instrumentation were evaluated for the ability to separate and determine molar mass (MM) of starch polymers, respectively. The determination of M_w by MALLS is necessary because the M_w of many cereal starches exceeds the available molecular standards by one or two orders of magnitude. The M_w depends on the method of calculation. The M_w (Berry method) of starch from waxy corn was 2.27 × 10⁸ Da, waxy rice 8.9 × 10⁷ Da, cassava 5.7 × 10⁷ Da, Hylon V 2.7 × 10⁷ Da, Hylon VII 4.8 × 10⁶ Da, and potato amylose 1.9 × 10⁵ Da. Recovery dropped dramatically for molecules with root mean square radii >200 nm.     

Relationships among the composition and physicochemical properties of starches with the characteristics of their films

The physical, molecular, and functional properties of corn, cassava, and yam starches were related to the film properties of these starches. Corn, cassava, and yam starches contained 25%, 19%, and 30% amylose, respectively. Amylose from yam starch showed the smallest molecular weight among the starches and amylopectin from corn starch the smallest molecular weight. Cassava starch presented a higher amylopectin content, and its gels and films were less strong, more transparent, and more flexible than corn and yam films. Plasticized films of the three starches were more flexible, with a higher strain and lower stress at break when the glycerol content increased. Unplasticized films were brittle and had water vapor permeability values ranging from 6.75 x 10(-10) to 8.33 x 10(-10) g m(-1) s(-1) Pa(-1). These values decreased when the glycerol content reached 20 g/100 g of starch because a more compact structure was formed. Then, at a glycerol content of 40 g/100 g of starch, the WVP increased because the film matrixes became less dense.     

木薯AGPase酶活性及其同工酶位点检测

本文以3个亲缘关系较远的高产木薯品种（SC124,SC8和Arg7）为材料,对块根发育过程中的AGPase活性和淀粉含量进行了测定,并利用AGPase同工酶电泳技术（非变性聚丙烯酰胺凝胶电泳）对AGPase酶活性较高时期同工酶位点进行检测,旨在分析木薯块根AGPase同工酶位点与AGPase酶活性和淀粉积累的关系。结果表明：木薯块根发育过程中AGPase酶活性和淀粉含量呈极显著正相关（R=0．80212）,Arg7的AGPase酶活性和淀粉含量在块根发育都显著高于SC124和SC8。同工酶分析表明,木薯至少有6个同工酶位点（AGPa,AGPb,AGPc,AGPd,AGPe和AGPf）,且品种间具有多态性。其中,3个品种共有的4个同工酶位点分别为AGPa、AGPc、AGPd和AGPf,AGPe位点只在Arg7中出现,初步判断同工酶位点AGPe可能与其较高的酶活性和淀粉含量有关。本研究为系统研究木薯品种间AGPase酶活性与淀粉积累的关系提供了参考。     

Physicochemical Properties of Pueraria Root Starches and Their Effect on the Improvement of Buckwheat Noodle Quality

The physicochemical properties of starches from cultivated Pueraria thomsonii Benth were examined and compared with those of P. lobata (Willd.) Ohwi and other root starches, and the effect of pueraria root starches on the improvement of buckwheat noodle quality was investigated. The total content of isoflavones in P. thomsonii root starches was higher than in P. lobata root starches, and the size and uniformity of those particles displayed a significant difference. The gel stabilities of pueraria root starches were similar and more favorable than those of potato starch and sweet potato starch. For the amylose molecular properties of pueraria root starches, the λ_max and blue value index were higher than those of the potato starch and the sweet potato starch, whereas the amylose content and degree of polymerization were much lower in comparison. However, amylopectin branch lengths of pueraria root starches were shorter. Thus, pueraria root starches could improve the quality of buckwheat noodles and enhance their nutritional function. Therefore, pueraria root starches may be regarded as raw materials that influence the quality of buckwheat noodles.     

Effect of Starch Granule Size Distribution and Elevated Amylose Content on Durum Dough Rheology and Spaghetti Cooking Quality

To obtain an indication of the effect of increasing the starch amylose content above normal levels (27–74%) and increasing the percentage of B‐type starch granules (11–60%) on durum dough properties and the quality of the spaghetti derived from these doughs, the reconstitution approach was used. Reconstituted flours were prepared from a common Wollaroi gluten, solubles and tailings fraction combined with starches containing varying B‐granule contents, or with starches from maize with varying amylose content. An increased B‐granule content increased farinograph water absorption. Cooked spaghetti firmness was highest with B‐type granules at 32–44% (volume percentage basis), which is ≈10–15% higher than normally found in durum starch. Increasing the amylose content in the starch caused the dough to be more extensible, increased spaghetti firmness, and decreased water absorption with optimum quality of amylose at 32–44%. The information indicates there would be benefit in producing durum wheats with slightly elevated B‐granule and amylose contents.     

Photodegradation of cassava and corn starches

The baking expansion properties of sour cassava starch (Polvilho azedo) are attributable to photochemical starch degradation induced by heterolactic fermentation after sun-drying. This study investigated the effects of UV irradiation on the different structural levels of cassava starch as compared to those of corn starch and dextrins. Photosensitive compounds excited at 360 and 290 nm in cassava starch were photodegraded when starch was exposed to sunlight or 360 nm irradiation. UV irradiation depolymerized cassava and corn starches, inducing modifications due, at least in part, to a mechanism involving free radicals. Lactic acid was also photodegraded. Photodegradation induced by UV absorption could have been due to fluorescent chromophores found in starches and nonfluorescent chromophores present in glucosidic units.     

Sources of Variation for Starch Gelatinization,Pasting, and Gelation Properties in Wheat

The starch of wheat (Triticum aestivum L.) flour affects food product quality due to the temperature-dependent interactions of starch with water during gelatinization, pasting, and gelation. The objective of this study was to determine the fundamental basis of variation in gelatinization, pasting, and gelation of prime starch derived from seven different wheat cultivars: Kanto 107, which is a partial waxy mutant line, and six near-isogenic lines (NILs) differing in hardness. Complete pasting curves with extended 16-min hold at 93°C were obtained using the Rapid Visco Analyser (RVA). Apparent amylose content ranged from 17.5 to 23.5%; total amylose content ranged from 22.8 to 28.2%. Starches exhibited significant variation in onset of gelatinization. However, none of the parameters measured consistently correlated with onset or other RVA curve parameters that preceded peak paste viscosity. Peak paste viscosity varied from 190 to 323 RVA units (RVU). Higher peak, greater breakdown, lower final viscosity, negative setback, and less total setback were associated with lower apparent and total amylose contents. Each 1% reduction in apparent or total amylose content corresponded to an increase in peak viscosity of about 22 and 25 RVU, respectively, at 12% starch concentration. Of the seven U.S. cultivars, the lower amylose cultivars Penawawa and Klasic were missing the granule-bound starch synthase (GBSS; ADPglucose starch glycosyl transferase, EC 2.4.4.21) protein associated with the Waxy gene locus on chromosome 4A (Wx-B1 locus). Kanto 107 was confirmed as missing both the 7A and 4A waxy proteins (Wx-A1 and Wx-B1 loci). The hardness NIL also were shown to be null at the 4A locus. Apparent and total amylose contents of prime starch generally corresponded well to the number of GBSS proteins; although the hardness NIL tended to have somewhat higher amylose contents than did the other GBSS 4A nulls. We concluded that reduced quantity of starch amylose due to decreased GBSS profoundly affects starch gelatinization, pasting, and gelation properties.     

不同类型玉米籽粒淀粉积累、相关酶活及基因表达差异分析

为探究不同类型玉米淀粉形成机理,对普通玉米、甜玉米、糯玉米淀粉积累、相关酶活及基因表达进行测定,分析不同类型玉米淀粉积累、相关酶活及基因表达之间的差异及相互关系。结果表明,不同类型玉米总淀粉和直链淀粉百分含量为:普通玉米>糯玉米>甜玉米,支链淀粉百分含量为:糯玉米>普通玉米>甜玉米,灌浆期间总淀粉和直链淀粉含量3个玉米类型间差异显著;灌浆期间,普通玉米各淀粉合成相关酶活性最高,甜玉米淀粉合成相关酶活性最低,糯玉米则介于普通玉米和甜玉米之间,但其GBSS酶活性很小。灌浆期间3个类型玉米除GBSS酶活性差异不显著外,其他淀粉合成相关酶活性差异显著;普通玉米淀粉合成相关基因表达量总体均高于甜玉米和糯玉米,甜玉米和糯玉米相关突变基因仍存在表达。表明不同类型玉米淀粉含量和组成上差异明显;普通玉米淀粉的形成需要淀粉合成相关酶相互作用,淀粉合成相关酶活性的缺失会改变淀粉组成;不同类型玉米淀粉合成相关基因表达差异显著,但都存在转录活性。对普通玉米进行相关性分析同时发现,淀粉的合成不仅受到转录调控,还受到转录后调控,淀粉的合成是淀粉合成各酶之间相互协调的结果。     

Soft Wheat Starch Pasting Behavior in Relation to A‐ and B‐type Granule Content and Composition

Flours of two soft wheat cultivars were fractionated into native, prime, tailing, A‐, and B‐type starch fractions. Starch fractions of each cultivar were characterized with respect to A/B‐type granule ratio, amylose content, phosphorus level (lysophospholipid), and pasting properties to investigate factors related to wheat starch pasting behavior. While both cultivars exhibited similar starch characteristics, a range of A‐type (5.7– 97.9%, db) and B‐type granule (2.1–94.3%, db) contents were observed across the five starch fractions. Though starch fractions displayed only subtle mean differences (<1%) in total amylose, they exhibited a range of mean phosphorus (446–540 μg/g), apparent amylose (18.7–23%), and lipid‐complexed amylose (2.8–7.5%) values, which were significantly correlated with their respective A‐ and B‐type granule contents. A‐type (compared with B‐type) granules exhibited lower levels of phosphorus, lipid‐complexed amylose, and apparent amylose, though variability for the latter was primarily attributed to starch lipid content. While starch phosphorus and lipid‐complexed amylose contents exhibited negative correlation with fraction pasting attributes, they did not adequately account for starch fraction pasting behavior, which was best explained by the A/B‐type granule ratio. Fraction A‐type granule content was positively correlated with starch pasting attributes, which might suggest that granule size itself could contribute to wheat starch pasting behavior.