The influence of growth stages on cassava tuberous root rot in different ecological environments

A multilocational field experiment to determine the influence of cassava growth and development on tuberous root rots in Onne and Sabongidda-Ora humid forests and in the Ibadan derived savanna of Nigeria was carried out from June 2003 to November 2005. Healthy cassava stems of TME-1 (local), TMS 30572 and TMS 4 (2) 1425 (both improved) were planted in a randomised complete block design with three replications. The experiment was repeated and the data collected were statistically analysed using the general linear model statistical procedures with the SAS system for Windows. Experimental result showed tuberous root rot incidence and severity to be pronounced at 12 months after planting in Onne and Sabongidda-Ora humid forests but less at the Ibadan derived savanna. Local TME-1 variety had the highest root rot incidence of 53% and severity of 21% while improved TMS 4(2)1425 variety had the least root rot incidence and severity at 0% each.     

Regeneration of cassava plants via shoot organogenesis

A novel regeneration system based on direct shoot organogenesis is described for cassava. Plants could be regenerated at high frequency by inducing shoot primordia on explants derived from cotyledons of cassava somatic embryos. After a passage on elongation medium, the regenerated shoots were easily rooted in hormone-free medium and could be successfully transplanted to soil. Using the shoot-organogenesis-based regeneration method, up to eight transplantable plantlets per explant could be regenerated. The system was optimised first for one cassava cultivar, and then its transferability to three other cultivars was demonstrated. This method widens the scope of in vitro regeneration modes of cassava, and is also compatible with Agrobacterium-mediated transformation. To develop an efficient system for production of somatic embryos for regeneration experiments, conditions for inducing primary and cycling somatic embryos were also studied, and highly efficient plant regeneration via germination of somatic embryos was achieved using maltose instead of sucrose in the culture medium, and combining paclobutrazol with 2,4-dichlorophenoxyacetic acid in the embryo induction medium. Received: 25 January 1997 / Revision received: 10 February 1997 / Accepted: 20 February 1997     

植物生长调节物质IP－1号对木薯产量及其生物性状的影响

１９９０和１９９１年在木薯（ＭａｎｉｈｏｔｅｓｃｕｌｅｎｔａＣｒａｎｔｚ）生长期以植物生长调节物质ＩＰ－１号０,２０,３０和４０ｐｐｍ进行叶面喷洒,结果表明：３０ｐｐｍ处理可使木薯块根产量平均增加５４．４４％,块根淀粉含量平均提高２０．８１％。单株最大薯重提高３１．５５％,块根数增加２１．１７％,块根长度增长１７．６２％,地上部鲜重增加３４．３６％,植株高度增加４．３６％,植株收获期保留青叶数增加１９．４２％,主茎直径增加６．２６％,块根直径增加２．５８％,叶片的叶绿素和蛋白质含量分别提高５．５７％和２５．９６％,叶片光合作用强度提高１５．８６％,而对主茎高度、主茎节数没有明显影响。     

木薯的组织培养和基因转化

木薯是重要的热带作物之一, 其块根富含淀粉, 是热带、亚热带地区近５ 亿人粮食的主要来源。简单介绍近年来运用生物技术改良木薯的研究进展。     

Development of waxy cassava with different Biological and physico-chemical characteristics of starches for industrial applications

The quality of cassava starch, an important trait in cassava breeding programs, determines its applications in various industries. For example, development of waxy (having a low level of amylose) cassava is in demand. Amylose is synthesized by granule-bound starch synthase I (GBSSI) in plants, and therefore, down-regulation of GBSSI expression in cassava might lead to reduced amylose content. We produced 63 transgenic cassava plant lines that express hair-pin dsRNAs homologous to the cassava GBSSI conserved region under the control of the vascular-specific promoter p54/1.0 from cassava (p54/1.0::GBSSI-RNAi) or cauliflower mosaic virus (CaMV) 35S (35S::GBSSI-RNAi). After the screening storage roots and starch granules from field-grown plants with iodine staining, the waxy phenotype was discovered: p54/1.0::GBSSI-RNAi line A8 and 35S::GBSSI-RNAi lines B9, B10, and B23. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that there was no detectable GBSSI protein in the starch granules of plants with the waxy phenotype. Further, the amylose content of transgenic starches was significantly reduced (<5%) compared with the level in starch granules from the wild-type (about 25%). The inner structure of the waxy starch granules differed from that of the untransformed ones, as revealed by transmission electron microscopy analysis as well as morphological changes in the iodine-starch complex. Endothermic enthalpy was reduced in waxy cassava starches, according to differential scanning calorimeter analysis. Except B9, all waxy starches displayed the A-type X-ray diffraction pattern. Amylogram patterns of the waxy cassava starches were analyzed using a rapid viscosity analyzer and found to have increased values for clarity, peak viscosity, gel breakdown, and swelling index. Setback, consistency, and solubility were notably reduced. Therefore, waxy cassava with novel starch in its storage roots was produced using the biotechnological approach, promoting its industrial utilization.     

How in vitro light affects growth and survival of ex vitro cassava

Cassava is one of the most important food crops in Africa. Meristem culture is an effective method of eliminating viruses and other systemic diseases spread through the vegetative propagation of stems. However, in semi‐arid conditions, survival of ex vitro plants in the field is often disappointing. When an increasing range of light regimes in vitro was provided, the fresh and dry masses more than doubled their values between 29 and 369 mmol s^?1 m^?2 PPFDs. Increases in numbers of senescent leaves and stem thickness were also recorded with increasing PPFD. However, PPFD above 101 mmol s^?1 m^?2 resulted in 30–70% reduction in plant survival, with the thin plants with the smallest fresh and dry masses being the ones with highest survival rates. High light and temperature levels in the greenhouse were also found to be critical for plant survival. It was also shown that transpirational loss from detached leaves and epicuticular wax deposits were not good indicators for predicting survival of ex vitro cassava plantlets during acclimatisation.     

影响根癌农杆菌介导的木薯遗传转化因素分析

采用根癌农杆菌介导的叶盘转化法,以我国南方地区主栽木薯品种—华南8号的胚状体子叶为受体,对影响木薯遗传转化效率的主要因素进行了分析。研究结果表明,在木薯的遗传转化中,选用GV3101作为浸染外植体的农杆菌菌株,将感染时间和共培养时间分别控制在30~45 min和3~4 d、菌液浓度（OD₆₀₀）采用0.45、并添加200 μmol·L^-1的乙酰丁香酮（AS）均可明显提高其转化效率,但若对外植体进行预培养反而会降低其转化效率。利用该体系从453块外植体中共转化获得10株抗性再生植株,经PCR和Southern杂交检测,有8株木薯的基因组中已整合进了外源基因glgC336,转化率为1.77%。     

Using microsatellites,isozymes and AFLPs to evaluate genetic diversity and redundancy in the cassava core collection and to assess the usefulness of DNA-based markers to maintain germplasm collections

The cassava core collection was selected to represent, with minimum repetitiveness, the potential genetic diversity of the crop. The core (630 accessions) was chosen from the base collection (over 5500 accessions) on the basis of diversity of origin (country and geographic), morphology, isozyme patterns and specific agronomic criteria. To asses the genetic diversity of the core, 521 accessions were typed with four microsatellite loci. Allele diversity and frequency, and size variance of dinucleotide repeats (Rst statistic) were estimated. Microsatellite allele numbers and frequencies varied among countries: Colombia and Brazil had the largest number of different alleles across all loci. Mexico also had a high number, ranking fifth after Peru, Costa Rica and Venezuela (which tied). Unique alleles were present in accessions from Brazil, Colombia, Guatemala, Venezuela and Paraguay. A small number (1.34%) of potential duplicates were identified through isozyme and AFLP profiles. Thus, the present results indicated that traditional markers have been highly effective at selecting unique genotypes for the core. Future selections of cassava germplasm sets can be aided by DNA-based markers to ensure genetically representative, non-redundant samples. This revised version was published online in June 2006 with corrections to the Cover Date.     

Production of stably transformed cassava plants via particle bombardment

 A novel protocol, based on biolistics and regeneration via organogenesis, was developed for genetic transformation of cassava (Manihot esculenta Crantz). The in vitro performance of cassava cultivars CMC40, MPer183 and MCol22 was evaluated, and the regeneration protocol was modified to improve shoot production from explants for transformation experiments. Somatic cotyledons were used as a target tissue in the transformation experiments using the Particle Inflow Gun and a plasmid containing the uidA gene in transient assays. The effect of different parameters for particle bombardment efficiency, including the amount of DNA used, the flying distance of the projectiles and the pre- and post-plasmolysis time of the target tissue, was evaluated and the conditions were partially optimised. Stably transformed cassava plants of cvs. MCol22 and TMS60444 were produced using the partially optimised conditions and two different vector constructs carrying the hpt gene as the selectable marker. The selection protocol was optimised further, and a rooting test was developed for screening the regenerants for antibiotic resistance to reduce the number of escapes obtained after primary selection. The production of stably transformed cassava lines and the expression of the transgenes was verified by Southern blot analysis and RT-PCR. Received: 10 December 1999 / Revision received: 12 January 2000 / Accepted: 7 February 2000     

The major economic field diseases of cowpea in the humid agro-ecologies of south-western Nigeria

Cowpea, which has now become an important protein source for the teeming populace of Nigerians, especially those living in the humid agro-ecological zones of south-western Nigeria, is severely attacked by diseases. The causal agents of these diseases find the environment more conducive for survival and hence induced disease conditions in the host plant, causing significant yield reduction. This paper reviews the present situation of the cowpea diseases in the humid forest agro-ecologies of south-western Nigeria.     

Occurrence of Fusarium andiyazi associated with sorghum in Nigeria

Investigations into fungi associated with sorghum grain in Nigeria indicate the occurrence of a newly described fungus, Fusarium andiyazi alongside F. nygamai. These fungi have earlier been reported as F. moniliforme. Our results highlight the need to re-evaluate Fusarium species associated with sorghum in Nigeria.     

Distinguishing the effects of light and temperature variations on the growth, development, multiplication potential and ex vitro survival rates of in vitro cassava

Dissemination of cassava tissue culture plantlets is difficult in the arid tropics due to low eux vitro survival rates. Increased in vitro light intensity has been reported to induce high ex vitro survival rates. The results from earlier experiments suggested that it would be worthwhile to analyse the separate effects of in vitro light and of temperature on the in vitro growth pattern as well as differentiate its relation to ex vitro survival. Accordingly, analysis of a range of in vitro light intensities from 0 to 369 μmol^?1 m^?2 photosynthetic photon flux density (PPFD) was conducted both with and without fans to control the heat. Temperature proved stable at low PPFD levels but increased above 204 μmol s^?1 m^?2 when no fans were used. Increased PPFD levels induced larger fresh and dry masses as well as stem thickness. PPFD levels affected the developmental index (senescent leaf numbers) in vitro when it rose above 204 μmol s^?1 m^?2 PPFD. Raised temperature ranges increased the multiplication index (node numbers) in vitro and ex vitro. It increased root number and leaf development (lobe anatomy). As in vitro temperatures of up to 40°C improve multiplication rates and PPFD levels above 101 μmol s^?1 m^?2 were detrimental for ex vitro survival (as low as 60%), it is suggested that simpler and less costly laboratories with low light levels and a wide range of temperature tolerance could be successfully established in the tropics for in vitro cleaning and rapidly multiplying crops like cassava.     

Large-scale genome-wide association study,using historical data,identifies conserved genetic architecture of cyanogenic glucoside content in cassava (Manihot esculenta Crantz) root

Manihot esculenta (cassava) is a root crop originating from South America that is a major staple in the tropics, including in marginal environments. This study focused on South American and African germplasm and investigated the genetic architecture of hydrogen cyanide (HCN), a major component of root quality. HCN, representing total cyanogenic glucosides, is a plant defense component against herbivory but is also toxic for human consumption. We genotyped 3354 landraces and modern breeding lines originating from 26 Brazilian states and 1389 individuals were phenotypically characterized across multi-year trials for HCN. All plant material was subjected to high-density genotyping using genotyping by sequencing. We performed genome-wide association mapping to characterize the genetic architecture and gene mapping of HCN. Field experiments revealed strong broad- and narrow-sense trait heritability (0.82 and 0.41, respectively). Two major loci were identified, encoding for an ATPase and a MATE protein, and contributing up to 7 and 30% of the HCN concentration in roots, respectively. We developed diagnostic markers for breeding applications, validated trait architecture consistency in African germplasm and investigated further evidence for the domestication of sweet and bitter cassava. Fine genomic characterization revealed: (i) the major role played by vacuolar transporters in regulating HCN content; (ii) the co-domestication of sweet and bitter cassava major alleles are dependent upon geographical zone; and (iii) the major loci allele for high HCN in M. esculenta Crantz seems to originate from its ancestor, M. esculenta subsp. flabellifolia. Taken together, these findings expand our insights into cyanogenic glucosides in cassava roots and its glycosylated derivatives in plants.     

Over-expression of hydroxynitrile lyase in transgenic cassava roots accelerates cyanogenesis and food detoxification

Cassava (Manihot esculenta, Crantz) roots are the primary source of calories for more than 500 million people, the majority of whom live in the developing countries of Africa. Cassava leaves and roots contain potentially toxic levels of cyanogenic glycosides. Consumption of residual cyanogens (linamarin or acetone cyanohydrin) in incompletely processed cassava roots can cause cyanide poisoning. Hydroxynitrile lyase (HNL), which catalyses the conversion of acetone cyanohydrin to cyanide, is expressed predominantly in the cell walls and laticifers of leaves. In contrast, roots have very low levels of HNL expression. We have over-expressed HNL in transgenic cassava plants under the control of a double 35S CaMV promoter. We show that HNL activity increased more than twofold in leaves and 13-fold in roots of transgenic plants relative to wild-type plants. Elevated HNL levels were correlated with substantially reduced acetone cyanohydrin levels and increased cyanide volatilization in processed or homogenized roots. Unlike acyanogenic cassava, transgenic plants over-expressing HNL in roots retain the herbivore deterrence of cyanogens while providing a safer food product.     

In vitro and greenhouse evaluations of Trichoderma isolates for biological control of potato wilt disease (Fusarium solani)

The biological efficacy of Trichoderma species may differ due to variations in ecosystems. This study was conducted to assess the biocontrol efficacy of some native Trichoderma isolates against Fusarium solani, an important causal agent of potato wilt disease under laboratory and greenhouse conditions at Shahrood Agricultural Research Centre, Shahrood, Iran, during 2006–2007. Fourteen isolates were collected among which eight showed promising ability in inhibiting the growth of the pathogen through dual culture and production of volatile and non-volatile inhibitors but T. brevicompactum (T1), T. longibrachiatum (T5) and T. asperellum (T2) were almost better than other isolates in inhibiting the mycelial growth of the pathogen in comparison to control in the above three tests (p ≤ 0.01). Isolates performing mycoparasitism under in vitro condition were evaluated against the disease in pot culture under greenhouse condition. In all treatments in which Trichoderma isolates + F. solani were involved lower disease incidence was noticed in comparison to Fusarium-infested control (p ≤ 0.05). Best disease control was observed in potted plants treated with F. solani + T. longibrachiatum (T5) with 6.25% disease incidence in comparison to Fusarium-infested control, in which disease incidence was observed to be 75%. Interaction of T. brevicompactum (T1) and F. solani also indicated good control of the disease by 12.50% of disease incidence.     

Genetic variability of Fusarium wilt pathogen isolates of chickpea (Cicer arietinum L.) assessed by molecular markers

Genetic variability among 43 isolates of Fusarium oxysporum f.sp. ciceri, the chickpea wilt pathogen, collected from nine states of India including the four well-characterized races of the pathogen were assessed using the molecular markers, RAPDs and AFLP. Principal coordinate analysis of the similarity index data generated from the molecular marker studies mostly gave three different clusters: Of these two clusters represented race-1 and race-2, and the third cluster consisted of race-3 and race-4 pathogen isolates. In RAPDs a fourth cluster was seen which did not go with any of the four races of the pathogen. The molecular markers established the distinctness of race-1 and race-2 pathogen isolates and the close similarity of pathogen isolates of race-3 with that of race-4. AFLP was found to be more informative as it differentiated more number of the pathogen isolates with the known races with minimum of outliers. The high levels of DNA polymorphism observed with the molecular markers suggest the rapid evolution of new recombinants of the pathogen in the chickpea growing fields. This revised version was published online in June 2006 with corrections to the Cover Date.     

木薯雌雄花分化形态结构观察及生理调控研究

本研究以木薯品种新选048为试验材料,观察和测定木薯花分化各时期的形态结构、内源激素及碳氮化合物含量,并开展木薯雌雄花分化的外源激素调控研究,揭示木薯花分化规律,初步探究木薯花分化的生理机制.试验结果表明:木薯从花芽分化开始到完成开花的全过程需要30 d,分化出的花序主要有全雄花序和雌少雄多花序两种类型,木薯性别分化前...     

Characterisation,differentiation and biochemical diversity of Fusarium solani and Fusarium proliferatum based on cellular fatty acid profiles

The biochemical relationships between Fusarium solani and Fusarium proliferatum isolates were investigated using fatty acid analysis. Cellular fatty acid composition showed that palmitic acid, stearic acid, oleic acid and linoleic acid were the most abundant fatty acids in these species and accounted for 93.88 and 94.02% of the fatty acid profiles in F. solani and F. proliferatum, respectively. The most predominant fatty acids were linoleic acid (37.44%) in F. solani and oleic acid (39.81%) in F. proliferatum. The fatty acid compositions of F. solani and F. proliferatum were significantly different (p?<?0.05) for most of the individual fatty acids. This study demonstrated that fatty acid profiles may be useful to characterise and differentiate F. solani and F. proliferatum isolates at the species level. Using fatty acid analysis, biochemical diversity was observed among isolates of these species. The dendrogramme revealed that F. solani and F. proliferatum formed two distinct clusters with a distance of 7.2. Isolates of each species were clustered with each other, having a Euclidean distance of 6 and 6.6 for F. solani and F. proliferatum, respectively.     

Effects of cytokinins and auxins on cassava shoot organogenesis and somatic embryogenesis from somatic embryo explants

Somatic embryos of cassava (Manihot esculenta Crantz) cultivar ‘Nanzhi 188’ were isolated and cut into fragments to be cultured on media with various cytokinins and auxins. Cytokinin induced of cassava shoot organogenesis, while auxin stimulated somatic embryogenesis. The effectiveness on organogenesis was different based on different cytokinins and in combination with auxins. Benzyladenine and thidiazuron stimulated more shoot organogenesis than kinetin and N-isopentenyladenine. This revised version was published online in June 2006 with corrections to the Cover Date.     

Effect of water deficits on transpiration, photosynthesis and leaf conductance in cassava

Transpiration, net photosynthesis and leaf conductance decreased when leaf water potential dropped below -0.30 MPa. Both transpiration and net photosynthesis rates were considerably reduced before the leaves were visibly wilted at -0.95 MPa. Consequently, visual symptoms are unlikely to provide a useful index for characterizing water deficits in cassava ( Manihot esculenta Crantz cv. Llanera). Decreases in net photosynthesis closely followed decreases in transpiration and this suggests that stomatal closure controls both processes.