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。     

Putting plant resistance traits on the map: a test of the idea that plants are better defended at lower latitudes

? It has long been believed that plant species from the tropics have higher levels of traits associated with resistance to herbivores than do species from higher latitudes. A meta-analysis recently showed that the published literature does not support this theory. However, the idea has never been tested using data gathered with consistent methods from a wide range of latitudes. ? We quantified the relationship between latitude and a broad range of chemical and physical traits across 301 species from 75 sites world-wide. ? Six putative resistance traits, including tannins, the concentration of lipids (an indicator of oils, waxes and resins), and leaf toughness were greater in high-latitude species. Six traits, including cyanide production and the presence of spines, were unrelated to latitude. Only ash content (an indicator of inorganic substances such as calcium oxalates and phytoliths) and the properties of species with delayed greening were higher in the tropics. ? Our results do not support the hypothesis that tropical plants have higher levels of resistance traits than do plants from higher latitudes. If anything, plants have higher resistance toward the poles. The greater resistance traits of high-latitude species might be explained by the greater cost of losing a given amount of leaf tissue in low-productivity environments.     

Phylogenetic patterns in the genus Manihot (Euphorbiaceae) inferred from analyses of nuclear and chloroplast DNA regions

From a phylogenetic perspective, the genus Manihot can be considered as an orphan group of plants, and the scientific knowledge acquired has been mainly related to cassava, one of the most important crops in poor tropical countries. The goal of the majority of evolutionary studies in the genus has been to decipher the domestication process and identify the closest relatives of cassava. Few investigations have focused on wild Manihot species, and the phylogeny of the genus is still unclear. In this study the DNA sequence variation from two chloroplast regions, the nuclear DNA gene G3pdh and two nuclear sequences derived from the 3'-end of two cassava ESTs, were used in order to infer the phylogenetic relationships among a subset of wild Manihot species, including two species from Cnidoscolus as out-groups. Maximum parsimony and Bayesian analyses were conducted for each data set and for a combined matrix due to the low variation of each region when analyzed independently. A penalized likelihood analysis of the chloroplast region trnL-trnF, calibrated with various age estimates for genera in the Euphorbiaceae extracted from the literature was used to determine the ages of origin and diversification of the genus. The two Mesoamerican species sampled form a well-defined clade. The South American species can be grouped into clades of varying size, but the relationships amongst them cannot be established with the data available. The age of the crown node of Manihot was estimated at 6.6 million years ago. Manihot esculenta varieties do not form a monophyletic group that is consistent with the possibility of multiple introgressions of genes from other wild species. The low levels of variation observed in the DNA regions sampled suggest a recent and explosive diversification of the genus, which is confirmed by our age estimates.     

引种保护植物对西双版纳极端冷年的春季物候响应

气温被普遍认为是春季物候期最主要的控制因子之一,然而低温对植物物候的影响效应一直都存在不同的观点。西双版纳由于地处热带地区的北缘,其气温相对于赤道附近的热带地区较低。自1959年以来,西双版纳热带植物园引入了来自世界各个热带地区的4万余种植物进行保护,之前的研究证明西双版纳的低温对这些引种植物的生长有很大影响。因此,1974年西双版纳出现的极端低温势必对引种植物造成极大威胁,同时也是对这些植物低温适应能力的一个考验。通过对比43种引种植物物候期（生长抽梢期与开花期）在1974年与常年的差异情况,分析不同来源（热带亚洲、热带美洲与热带非洲）引种植物对西双版纳低温的适应性。结果表明,经历西双版纳1974年初的极端低温之后,使81%的引种植物生长抽梢期提前,同时也造成35%的引种植物在该年没有开花;而植物生长抽梢提前的主要原因则是极端低温以及低温过后气温迅速回升。引种植物均能顺利度过1974年的最冷时期,并出现生长抽梢物候,这意味着引种植物在经历极端低温之后都能够进行正常的生长活动,但极端低温对引种植物繁殖活动的不利影响大于其对生长活动的影响;引种植物对西双版纳极端低温的适应能力由大到小顺序依次为：亚洲来源植物〉美洲来源植物〉非洲来源植物。因此在迁地保护植物的选择过程中,应多选择亚洲热带植物,其次为美洲热带植物,而对非洲热带植物的引入则需谨慎考察。     

引种保护植物对西双版纳极端冷年的春季物候响应

气温被普遍认为是春季物候期最主要的控制因子之一, 然而低温对植物物候的影响效应一直都存在不同的观点。西双版纳由于地处热带地区的北缘, 其气温相对于赤道附近的热带地区较低。自1959年以来, 西双版纳热带植物园引入了来自世界各个热带地区的4万余种植物进行保护, 之前的研究证明西双版纳的低温对这些引种植物的生长有很大影响。因此,1974年西双版纳出现的极端低温势必对引种植物造成极大威胁, 同时也是对这些植物低温适应能力的一个考验。通过对比43种引种植物物候期(生长抽梢期与开花期)在1974年与常年的差异情况, 分析不同来源(热带亚洲、热带美洲与热带非洲)引种植物对西双版纳低温的适应性。结果表明, 经历西双版纳1974年初的极端低温之后, 使81%的引种植物生长抽梢期提前, 同时也造成35%的引种植物在该年没有开花; 而植物生长抽梢提前的主要原因则是极端低温以及低温过后气温迅速回升。引种植物均能顺利度过1974年的最冷时期, 并出现生长抽梢物候, 这意味着引种植物在经历极端低温之后都能够进行正常的生长活动, 但极端低温对引种植物繁殖活动的不利影响大于其对生长活动的影响; 引种植物对西双版纳极端低温的适应能力由大到小顺序依次为: 亚洲来源植物>美洲来源植物>非洲来源植物。因此在迁地保护植物的选择过程中, 应多选择亚洲热带植物, 其次为美洲热带植物, 而对非洲热带植物的引入则需谨慎考察。     

Cassava genetic transformation and its application in breeding

As a major source of food, cassava (Manihot esculenta Crantz) is an important root crop in the tropics and subtropics of Africa and Latin America, and serves as raw material for the production of starches and bioethanol in tropical Asia. Cassava improvement through genetic engineering not only overcomes the high heterozygosity and serious trait separation that occurs in its traditional breeding, but also quickly achieves improved target traits. Since the first report on genetic transformation in cassava in 1996, the technology has gradually matured over almost 15 years of development and has overcome cassava genotype constraints, changing from mode cultivars to farmer-preferred ones. Significant progress has been made in terms of an increased resistance to pests and diseases, biofortification, and improved starch quality, building on the fundamental knowledge and technologies related to planting, nutrition, and the processing of this important food crop that has often been neglected. Therefore, cassava has great potential in food security and bioenergy development worldwide.     

Intermediary metabolic changes in rabbits administered linamarin or potassium cyanide

Since cassava (Manihot esculenta Crantz) is a staple food item of several million people in the tropics, its toxicity cannot be underestimated. Therefore an attempt has been made to understand the metabolic changes caused by the chronic administration of sublethal doses linamarin, the principal cyanoglucoside of cassava, to rabbits. A significant rise in lactic acid and total cholesterol in liver and brain and a highly significant depletion of phospholipids of brain tissue was observed. There were also significant variations in the lactate dehydrogenase isoenzyme pattern of treated animals as compared with the control rabbits. The findings suggest that some of the biological effects of linamarin are similar to those of free cyanide.     

Bioreduction of aldehydes and ketones using Manihot species

Biocatalysis constitutes an important tool in organic synthesis, especially for the preparation of chiral molecules of biological interest. A series of aliphatic and aromatic aldehydes and two ketones were reduced using plant cell preparations from Manihot esculenta and Manihot dulcis roots. The reduced products were typically obtained in excellent yields (80-96%), and with excellent enantiomeric excess (94-98%), except for vanillin. Esters, a nitrile, and an amide were also examined, but were not reduced. Preliminary conversion rate studies are reported. This is the first attempt to perform the biotransformation of carbonyl compounds using Manihot species.     

Clinal variation in Drosophila serrata for stress resistance and body size

Clines for size and stress resistance traits have been described for several Drosophila species and replicable clines across different species may indicate climatic selection. Here we consider clines in stress resistance traits in an Australian endemic species, D. serrata, by comparing levels of variation within and among isofemale lines initiated with flies collected from the eastern coast of Australia. We also consider clinical variation in chill coma recovery, a trait that has recently been shown to exhibit high levels of variation among Drosophila species. Patterns were compared with those in the cosmopolitan species D. melanogaster from the same area. Both desiccation and starvation resistance showed no clinical pattern despite heritable variation among isofemale lines. In contrast chill coma resistance exhibited a linear cline in the anticipated direction, resistance increasing with latitude. Body size was measured as wing length and body weight. Both traits showed geographic variation and strong non-linear clines with a sharp reduction in size in the tropics. These results are discussed in the context of climatic selection and evolutionary processes limiting species borders.     

A comparative anatomical study in cassava diploid and tetraploid hybrids

Cassava, Manihot esculenta Crantz, is the most important crop for poor people in the tropics and subtropics. It is a woody plant scarcely studied from the anatomical viewpoint. Information on its anatomy is needed for use in plant breeding programs. An interspecific hybrid between cassava and M. oligantha Pax was obtained and artificially polyploidized by the second author. Hand cross sections were applied to the stem, colored by safranin and alcian blue, and mounted in synthetic resin. Tetraploid type showed a larger portion of medulla and absence of a growth ring. Vascular tissues in tetraploid were larger too. Both of these types had similar structure of vessel elements and articulated laticifers. Distribution of different tissues in the two types refers to more resistance to drought in the tetraploid type than in the diploid one.     

The response of five tropical dicotyledon species to solar ultraviolet-B radiation

Tropical regions currently receive the highest levels of global solar ultraviolet-B radiation (UV-B, 280–320 nm) even without ozone depletion. The influence of natural, present-day UV-B irradiance in the tropics was examined for five tropical species including three native rain forest tree species (Cecropia obtusifolia, Tetragastris panamensis, Calophyilum longifolium) and two economically important species (Swietenia macrophylla, Manihot esculenta). Solar UV-B radiation conditions in a small clearing on Barro Colorado Island, Panama (9° N), were obtained using either a UV-B-excluding plastic film or a film that transmits most of the solar UV-B. Significant differences between UV-B-excluded and near-ambient UV-B plants were often exhibited as increased foliar UV-B absorbing compounds and, in several cases, as reduced plant height with exposure to solar UV-B. Increases in leaf mass per area and reductions in leaf blade length under solar UV-B occurred less frequently. Biomass and photosystem II function using chlorophyll a fluorescence were generally unaffected. The results of this study provide evidence that tropical vegetation, including native rain forest species, responds to the present level of natural solar UV-B radiation. This suggests that even minor ozone depletion in the tropics may have biological implications.     

Characterization of microsatellite markers in cassava based on microsatellite-AFLP technique

We developed molecular markers for cassava based on the microsatellite-amplified fragment length polymorphism (M-AFLP) technique. Twenty primer pairs were developed and used for the analysis of 48 samples of Manihot species, consisting of M. esculenta (33), M. esculenta ssp flabellifolia (3), M. chlorosticta (3), M. carthaginensis (3), M. filamentosa (3), and M. tristis (3). Nine microsatellite loci that were polymorphic among these Manihot species were identified, giving 32 polymorphic alleles and from two to seven alleles per locus. Unbiased and direct count heterozygosity varied from 0.0233 to 0.7924 and 0.0000 to 0.7083, respectively. There was significant deviation (P < 0.05) from Hardy-Weinberg equilibrium at five loci. Genotypic data from the Manihot species were subjected to genetic diversity analysis. We found that M. chlorosticta and M. esculenta ssp flabellifolia were the closest populations, while M. filamentosa and M. esculenta ssp flabellifolia were the most divergent. Considering within M. esculenta, the samples from Nigeria and Fiji were the most closely related, while those from Venezuela and of unknown origin were the most divergent. We conclude that the M-AFLP technique is an effective method for generating microsatellite markers that are useful for genetic diversity analysis in Manihot species.     

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.     

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.     

Aflatoxins of Aspergillus flavus Grown on Cassava

Aspergillus flavus Link ex Fries was grown on cassava (Manihot utilissima) and Czapek-Dox media at 31±1°C and 90 per cent relative humidity for 8 weeks. Isolation and purification of the toxic and carcinogenic metabolic products of the mould by paper and thin layer chromatography are described. High concentrations of aflatoxin B1, B2, G1 and G2 were synthesized by the mould grown on cassava for 4 weeks. In addition to these four major toxic components, 9 other fluorescent materials were observed on the chromatograms of crude products from cassava. It is concluded that the high moisture, high polysaccharide and low nitrogen content of cassava constitute a favourable nutritional condition at high temperatures and high relative humidity for the growth of A. flavus and the synthesis of relatively high concentrations of aflatoxin. This conld represent a serious health hazard in the moist tropics.     

Mapping EST-derived SSRs and ESTs involved in resistance to bacterial blight in Manihot esculenta.

Cassava (Manihot esculenta Crantz) is a major root crop widely grown in the tropics. Cassava bacterial blight, caused by Xanthomonas axonopodis pv. manihotis (Xam), is an important disease in Latin America and Africa resulting in significant losses. The preferred control method is the use of resistant genotypes. Mapping expressed sequence tags (ESTs) and determining their co-localization with quantitative trait loci (QTLs) may give additional evidence of the role of the corresponding genes in resistance or defense. Twenty-one EST-derived simple sequence repeats (SSRs) were mapped in 16 linkage groups. ESTs showing similarities with candidate resistance genes or defense genes were also mapped using strategies such as restriction fragment length polymorphisms, cleaved amplified polymorphic sequences, and allele-specific primers. In total, 10 defense-related genes and 2 bacterial artificial chromosomes (BACs) containing resistance gene candidates (RGCs) were mapped in 11 linkage groups. Two new QTLs associated with resistance to Xam strains CIO121 and CIO151 were detected in linkage groups A and U, respectively. The QTL in linkage group U explained 61.6% of the phenotypic variance and was associated with an RGC-containing BAC. No correlation was found between the new EST-derived SSRs or other mapped ESTs and the new or previously reported QTLs.     

SNPs,SSRs and inferences on cassava’s origin

Despite its importance as a staple food throughout the tropics, the root crop cassava (it Manihot esculenta ssp. esculenta) has traditionally not been a major focus of research. One basic question about cassava that remained unresolved until recently concerns the crop’s origin. This paper describes analyses of SNPs (single nucleotide polymorphisms) and SSR (simple sequence repeat) variation as a means of tracing cassava’s evolutionary and geographical origins. Genetic diversity was examined in a sample of 20 cassava varieties that are representative of germplasm diversity within the crop, and in 212 individuals collected from wild populations of two closely related Manihot species. SNP and indel variation was examined in portions of two low copy nuclear genes, BglA and Hnl. Inferences from these genes were compared to those from previously examined loci, including the low copy nuclear gene G3pdh and 5 SSR loci. For all genes examined, SNPs and SSR alleles are shared between domesticated cassava and a specific geographical subset of wild Manihot populations, which suggests the following: (1) Cassava was likely domesticated from a single wild Manihot species, M. esculenta ssp. flabellifolia, rather than from multiple hybridizing species, as traditionally believed; and (2) the crop most likely originated in the southern Amazon basin.     

The Latitudinal Diversity Gradient Through Deep Time: Testing the “Age of the Tropics” Hypothesis Using Carboniferous Productidine Brachiopods

The latitudinal gradient in global diversity, in which the number of species decreases away from the tropics, is widely recognized and well-studied but no consensus exists as to its cause. The “Age of the Tropics” hypothesis argues that the tropics have more species because young, ecologically dominant, tropical clades have not had sufficient time to disperse and adapt to colder climates; given time, the tropics act as a diversity pump. This hypothesis is tested using Carboniferous productidine brachiopods, which at that time were a young, ecologically dominant clade that originated in the tropics. A database of geographic occurrences indicates that productidines did not manifest a pattern of dispersal away from the tropics through time. A phylogeny of one productidine clade demonstrates that many lineages contracted, rather than expanded, their ranges over time. The results suggest that the hypothesis that the tropics are a diversity pump cannot be generalized across time and taxa.     

Climate, niche conservatism, and the global bird diversity gradient

We tested the proposition that there are more species in the tropics because basal clades adapted to warm paleoclimates have been lost in regions now experiencing cool climates. Molecular phylogenies were used to classify species as "basal" and "derived" based on their family, and their richness patterns were contrasted. Path models also evaluated environmental predictors of richness patterns. As predicted, basal clades are more diverse in the lowland tropics, whereas derived clades are more diverse in the extratropics and high-altitude tropics. Seventy-four percent of the variation in bird richness was explained by environmental variables, but models differed for basal and derived groups. The overall gradient is described by the spatial pattern of basal clades, although there are differences in the Old and New Worlds. We conclude that in ecological time, the global richness gradient reflects birds' responses to climatic gradients, partially operating via plants. Over evolutionary time, the gradient primarily reflects the extirpation of species in older clades from parts of the world that have become cooler in the present. A strong secondary effect arises from dispersal of clades from centers of origin and subsequent radiations. Overall, the diversity gradient is well explained by niche conservatism and the "time-for-speciation" hypothesis.