能源木薯种质资源面临的问题与解决策略

本文从种质资源角度,探讨了如何通过木薯新品种引进及基因工程开展木薯的定向培育研究。对淀粉品质、抗逆境特别是抗低温能力及采后生理性变质涉及的分予机制与生物技术研究策略进行了的解析和讨论,为我国木薯生物技术发展提供参考。     

木薯分子育种中若干基本科学问题的思考与研究进展

木薯作为全球重要的薯类作物,既是热带地区粮食安全的保障,也是重要的淀粉工业原材料,保障其稳产、高产、优质一直是育种家不变的研究主题．当前,木薯品种选育正处在从杂交育种转向分子育种的发展阶段,深入解析木薯特有的经济性状和生物学特点是利用生物技术进行遗传改良的基础．不同于谷物类作物,木薯光合同化物的转运和库源分配的调控机制必有其独特之处;同时,储藏根的“库容”直接影响其产量．作为热带作物,了解木薯对低温和干旱的响应可为改良其抗逆境能力提供理论依据．不同于其他薯类作物,木薯储藏根特有的“采后生理性变质”问题亟待解决,其发生和调控机制的解析对延长木薯货架期意义重大．随着分子生物学的发展,针对上述各方面的研究日益深入,不仅激发了感兴趣的公众对这些问题的认知和思考,也激励了科研人员不断努力寻找解析相关机制的方法,为最终通过分子育种手段改良木薯提供思路和技术方案,揭开木薯的层层“面纱”,推动木薯分子育种的发展．     

植物分子育种技术在改良木薯种质方面的应用

木薯是热带、亚热带地区的重要粮食作物和经济作物。培育出生产性状更加优良的木薯品种,是推进木薯产业更快更好发展的重要基础。分子育种技术在培育优良木薯品种方面具有传统育种技术不可比拟的优势。该文介绍了近年来在提高植物抗寒性与病虫害抗性、降低氰苷含量、提高淀粉含量及组成、改变储存物种类、防止木薯收获后变质等方面的研究进展,以便在这些研究进展的基础上,利用植物分子育种技术加快获得具有抗逆能力提高、品质改良、产量增加、耐储藏等优良特性的木薯新品种。     

能源木薯高淀粉抗逆分子育种研究进展与展望

木薯(Manihot esculenta Crantz)是全球重要的粮食作物,也是我国非粮生物质能源发展的主要原材料。长期以来,传统杂交育种是木薯新品种培育的主要手段。随着全球生态的变化和木薯产业发展的推进,需要加速培育抗逆能力强、高淀粉的木薯新品种,因此,利用基因工程针对特定性状开展品种创新表现出巨大的潜力。随着组学技术的发展,在木薯基础研究领域,特别是针对储藏根发育、淀粉富集、逆境响应与调控等方面的研究逐步深入。强化木薯基础理论研究和发展应用技术,对推动能源木薯的产业化发展具有重要意义。     

木薯高、低粉品种贮藏根生长关键期的生理生化特性比较

通过研究木薯Manihot esculenta高粉和低粉品种贮藏根在发育关键期(形成–膨大期)主要生理生化性质的差异及其动态变化，为木薯栽培管理和育种改良提供理论基础。对高粉木薯品种‘辐选01’(FX01)和‘Kasetsart 50’(KU50)以及低粉品种‘华南124’(SC124)和‘9I’于种植后120、130、140、151和165 d五个时期分别取贮藏根样品，测定淀粉、可溶性糖、脱落酸(ABA)、保护酶、活性氧(ROS)、丙二醛(MDA)等生理生化指标并分析变化规律。结果表明，可溶性糖含量在贮藏根皮层中始终高于淀粉储藏区；高粉品种中总可溶性糖含量在贮藏根膨大期快速增加，且表现出较高的抗氧化酶活性和较低的ROS及MDA含量；ABA含量呈现出高粉品种比低粉品种低的趋势。木薯低粉和高粉品种中可溶性糖的分布与其转运后合成淀粉的规律相一致，高粉品种中淀粉的积累速率始终高于低粉品种，其较高的抗氧化酶活性和较低的ROS及MDA含量有利于木薯贮藏根中淀粉的积累。     

木薯/花生间作对根际土壤微生态的影响

为了探明木薯/花生间作在增产增收的同时对土壤微生态的影响,研究木薯和花生以不同行距(30 cm,40 cm,50 cm)间作的根际土壤养分、微生物及相关酶活性的变化。结果表明:木薯/花生间作可增加根际土壤细菌、真菌、放线菌及总微生物数量和微生物多样性,30 cm间作行距的木薯、花生根际土壤微生物总数量分别比单作木薯、花生增加了129.6%和101.1%;间作根际土壤碱解氮、有效磷、有效钾和有机质含量相比单作增加,50 cm间作花生的根际土壤碱解氮、有效磷、有效钾量比单作花生增加了145.9%~195.9%,30 cm间作木薯的根际土壤有效钾、有效磷含量分别比单作木薯增加了161.8%和187.9%;木薯/花生间作的根际土壤过氧化氢酶、酸性磷酸酶活性活性相比单作升高,间作土壤脲酶和蛋白酶活性相比单作降低,30 cm间作木薯的根际土壤过氧化氢酶活性比单作木薯增加了59.2%,50 cm间作花生的根际土壤蔗糖酶活性比单作花生增加了97.4%。可见,木薯/花生间作可改善根际土壤微生态坏境,且适宜的间作行距更利于土壤养分含量和微生物数量的增加。     

木薯块根淀粉高累积的相关酶的活性特征

木薯的价值在于其块根内高含量的淀粉。木薯淀粉不仅可以作为粮食,还可以作为原料制造乙醇燃料等。但是其块根中淀粉高效累积生化机制依然有待于解决。本研究比较性研究了大田生长的高、低淀粉木薯品种在4个生长关键时期的蔗糖和淀粉代谢相关的酶活性。结果表明,根中的淀粉合成途径的限速步骤酶腺苷二磷酸葡萄糖焦磷酸化酶(ADPGase)活性、叶部的蔗糖磷酸合成酶(SPS)活性与木薯块根内的淀粉含量呈正相关关系,而根淀粉降解酶活性则呈负相关关系。该研究表明通过提高根中的ADPGase和叶中的SPS活性,同时降低块根中的淀粉降解酶活性有可能可以提高木薯块根中的淀粉积累。     

木薯叶的成分

木薯(Manihot esculenta Crantz)是一种热带作物,木薯根富含淀粉,世界上约有5亿人把它作为主食。木薯的叶子长势茂盛并有极高的蛋白含量。尽管木薯叶可以在改善热带地区居民的食品营养结构方面起到重要的作用,但是,人们对叶的食用远不如对根的食用普遍,其原因在于木薯叶中含有氢氰酸。     

木薯己糖载体基因MeSTP7的克隆与功能验证

己糖载体属于单糖转运蛋白家族,具有转运单糖类物质的作用,广泛存在于植物体各组织中并参与植物体的生长与发育。为了研究木薯己糖载体转运单糖的能力与对木薯生长发育的影响,本试验利用木薯全基因组数据库,通过RT-PCR方法从木薯KU50中克隆得到了木薯己糖载体基因MeSTP7,酵母异源表达与亚细胞定位实验结果表明MeSTP7具有转运葡萄糖、果糖、甘露糖和半乳糖的能力,定位在细胞膜上;qRT-PCR结果表明MeSTP7在高淀粉木薯品种KU50和低淀粉木薯品种CAS36根发育过程中的表达量均呈现逐渐降低的趋势,MeSTP7在KU50和CAS36的叶片中的表达量均为最高。本研究克隆得到了木薯己糖载体基因MeSTP7,并验证了MeSTP7的功能,为木薯己糖载体的研究提供了理论参考。     

Accumulation of Hydroxycoumarins During Post-harvest Deterioration of Tuberous Roots of Cassava (Manihot esculenta Crantz)

The use of the root crop Cassava (Manihot esculenta Crantz)is constrained by its rapid deterioration after harvest. Chemicaland spectroscopic examination revealed the accumulation of fourhydroxycoumarins (esculin, esculetin, scopolin and scopoletin),compounds derived from the phenylpropanoid pathway, during thetime course of post-harvest deterioration. Fluorescence-microscopyrevealed their localization in the apoplast of the parenchyma.Scopoletin and scopolin showed the most dramatic increases inconcentration, peaking by day 2 after harvesting. A smallersecondary peak of scopoletin tended to be more pronounced incultivars showing lower susceptibility to deterioration. Evidencefor the metabolism of scopoletin to an insoluble coloured productby means of a peroxidase is presented. This product may be thecause of the discolouration of the vascular tissue during storage.Copyright 2000 Annals of Botany Company Cassava, hydroxycoumarins, Manihot esculenta, peroxidases, post-harvest physiological deterioration, wound response     

The Cassava Source–Sink project: opportunities and challenges for crop improvement by metabolic engineering

Cassava (Manihot esculenta Crantz) is one of the important staple foods in Sub‐Saharan Africa. It produces starchy storage roots that provide food and income for several hundred million people, mainly in tropical agriculture zones. Increasing cassava storage root and starch yield is one of the major breeding targets with respect to securing the future food supply for the growing population of Sub‐Saharan Africa. The Cassava Source–Sink (CASS) project aims to increase cassava storage root and starch yield by strategically integrating approaches from different disciplines. We present our perspective and progress on cassava as an applied research organism and provide insight into the CASS strategy, which can serve as a blueprint for the improvement of other root and tuber crops. Extensive profiling of different field‐grown cassava genotypes generates information for leaf, phloem, and root metabolic and physiological processes that are relevant for biotechnological improvements. A multi‐national pipeline for genetic engineering of cassava plants covers all steps from gene discovery, cloning, transformation, molecular and biochemical characterization, confined field trials, and phenotyping of the seasonal dynamics of shoot traits under field conditions. Together, the CASS project generates comprehensive data to facilitate conventional breeding strategies for high‐yielding cassava genotypes. It also builds the foundation for genome‐scale metabolic modelling aiming to predict targets and bottlenecks in metabolic pathways. This information is used to engineer cassava genotypes with improved source–sink relations and increased yield potential.     

Cassava: an appraisal of its phytochemistry and its biotechnological prospects

The present state of knowledge of the phytochemistry of small molecules isolated from the roots and leaves of cassava, Manihot esculenta Crantz (Euphorbiaceae), is reviewed. Cassava roots are an important source of dietary and industrial carbohydrates, mainly eaten as a source of starch, forming the staple food to over 500 million; additionally, the roots have value as a raw material for industrial starch production and for animal feed giving the crop high economic value, but it suffers markedly from post-harvest physiological deterioration (PPD). The hydroxycoumarins scopoletin and its glucoside scopolin as well as trace quantities of esculetin and its glucoside esculin are identified from cassava roots during PPD. The biotechnological prospects for cassava are also reviewed including a critical appraisal of transgenic approaches for crop improvement, together with its use for bioethanol production, due to cassava's efficient ability to fix carbon dioxide into carbohydrate.     

Comparative Transcriptomic Analysis of Storage Roots in Cassava During Postharvest Physiological Deterioration

Plant Molecular Biology Reporter - Cassava is an important starchy and food crop; however, the commercial value of cassava is seriously constrained by postharvest physiological deterioration (PPD)....     

Study of the Early Events Leading to Cassava Root Postharvest Deterioration

Cassava (Manihot esculenta Crantz) roots, the fourth most important food crop of the world, is the major carbohydrate source for more than 600 million people in Africa, parts of Latin America, Oceania, and Asia. Besides being a rich source of starch (～80% of root), the root is also rich in vitamin C, some carotenoids, calcium, and potassium. Upon harvest, roots begin a process of physiological decay within 24–36 h called postharvest physiological deterioration or PPD. The early events leading to PPD are not known. Research to date concerning the study of PPD has mostly focused on the signaling events several hours after harvest. Upon examination of physiological and biochemical changes occurring 3 or 4 h after cassava root detachment, changes in the nature and type of volatile compounds emitted, secondary metabolites accumulated, and changes in the expression of key genes in reactive oxygen species (ROS) turnover were observed along with a correspondent increase in tissue cytoplasmic singlet oxygen presence using radical-specific fluorescent imaging of tissue samples. It is likely that these findings have significant implications to help us understand and assist in dissection of the early events leading to the postharvest deterioration of cassava root.     

Cellulose and Lignin Contents are Negatively Correlated with Starch Accumulation,and Their Correlation Characteristics Vary Across Cassava Varieties

Journal of Plant Growth Regulation - Cassava storage roots contain large amounts of starch and low amounts of cellulose and lignin. However, the relationship between lignification with cellulose...     

An EST resource for cassava and other species of Euphorbiaceae

Cassava (Manihot esculenta) is a major food staple for nearly 600 million people in Africa, Asia, and Latin America. Major losses in yield result from biotic and abiotic stresses that include diseases such as Cassava Mosaic Disease (CMD) and Cassava Bacterial Blight (CBB), drought, and acid soils. Additional losses also occur from deterioration during the post-harvest storage of roots. To help cassava breeders overcome these obstacles, the scientific community has turned to modern genomics approaches to identify key genetic characteristics associated with resistance to these yield-limiting factors. One approach for developing a genomics program requires the development of ESTs (expressed sequence tags). To date, nearly 23000 ESTs have been developed from various cassava tissues, and genotypes. Preliminary analysis indicates existing EST resources contain at least 6000–7000 unigenes. Data presented in this report indicate that the cassava ESTs will be a valuable resource for the study of genetic diversity, stress resistance, and growth and development, not only in cassava, but also other members of the Euphorbiaceae family.     

In vitro cultured primary roots derived from stem segments of cassava (Manihot esculenta) can behave like storage organs

BACKGROUND AND AIMS: Cassava (Manihot esculenta) has three adventitious root types: primary and secondary fibrous roots, and storage roots. Different adventitious root types can also regenerate from in vitro cultured segments. The aim of this study was to investigate aspects of in vitro production of storage roots. METHODS: Morphological and anatomical analyses were performed to identify and differentiate each root type. Twenty-nine clones were assayed to determine the effect of genotype on the capacity to form storage roots in vitro. The effects of cytokinins and auxins on the formation of storage roots in vitro were also examined. KEY RESULTS: Primary roots formed in vitro and in vivo had similar tissue kinds; however, storage roots formed in vitro exhibited physiological specialization for storing starch. The only consistent diagnostic feature between secondary fibrous and storage roots was their functional differentiation. Anatomical analysis of the storage roots formed in vitro showed that radial expansion as a consequence of massive proliferation and enlargement of parenchymatous cells occurred in the middle cortex, but not from cambial activity as in roots formed in vivo. Cortical expansion could be related to dilatation growth favoured by hormone treatments. Starch deposition of storage roots formed in vitro was confined to cortical tissue and occurred earlier than in storage roots formed in vivo. Auxin and cytokinin supplementation were absolutely required for in vitro storage root regeneration; these roots were not able to develop secondary growth, but formed a tissue competent for starch storing. MS medium with 5 % sucrose plus 0.54 microM 1-naphthaleneacetic acid and 0.44 microM 6-benzylaminopurine was one of the most effective in stimulating the storage root formation. Genotypes differed significantly in their capacity to produce storage roots in vitro. Storage root formation was considerably affected by the segment's primary position and strongly influenced by hormone treatments. CONCLUSIONS: The storage root formation system reported here is a first approach to develop a tuberization model, and additional efforts are required to improve it. Although it was not possible to achieve root secondary growth, after this work it will be feasible to advance in some aspects of in vitro cassava tuberization.