甘蔗基因克隆研究进展

甘蔗是重要的糖料和能源作物,在我国,其糖产量占全国食糖产量的94%。甘蔗是高光合效率C4作物,其蔗糖含量是所有植物中最高的,其基因组中蕴藏着大量重要基因。因此,甘蔗基因克隆对甘蔗品种定向改良具有重要意义。从蔗糖代谢、生长发育和抗逆性三个方面全面总结甘蔗基因克隆的研究进展,并提出今后研究的建议,以期为今后甘蔗基因研究提供参考。     

Sugarcane (Saccharum X officinarum): A Reference Study for the Regulation of Genetically Modified Cultivars in Brazil

Global interest in sugarcane has increased significantly in recent years due to its economic impact on sustainable energy production. Sugarcane breeding and better agronomic practices have contributed to a huge increase in sugarcane yield in the last 30?years. Additional increases in sugarcane yield are expected to result from the use of biotechnology tools in the near future. Genetically modified (GM) sugarcane that incorporates genes to increase resistance to biotic and abiotic stresses could play a major role in achieving this goal. However, to bring GM sugarcane to the market, it is necessary to follow a regulatory process that will evaluate the environmental and health impacts of this crop. The regulatory review process is usually accomplished through a comparison of the biology and composition of the GM cultivar and a non-GM counterpart. This review intends to provide information on non-GM sugarcane biology, genetics, breeding, agronomic management, processing, products and byproducts, as well as the current technologies used to develop GM sugarcane, with the aim of assisting regulators in the decision-making process regarding the commercial release of GM sugarcane cultivars.     

Role of oviposition preference in an invasive crambid impacting two graminaceous host crops

Oviposition preference studies of the Mexican rice borer, Eoreuma loftini (Dyar), on sugarcane, Saccharum spp., and rice, Oryza sativa L., showed that drought stressed sugarcane was 1.8-fold more attractive based on egg masses/plant than well watered sugarcane. The E. loftini susceptible sugarcane cultivar LCP 85-384 was 1.6-fold more attractive than HoCP 85-845 based on numbers of eggs per egg mass. Egg masses were 9.2-fold more abundant and 2.3-fold larger on sugarcane than on rice. Rice, however, was preferred to sugarcane on a plant biomass basis. Oviposition on sugarcane occurred exclusively on dry leaf material, which increased under drought stress. Egg masses per plant increased on drought stressed sugarcane and were correlated with several foliar free amino acids essential for insect growth and development. The more resistant (based on injury) but more attractive (based on oviposition) rice cultivar XL8 had higher levels of several free amino acids than the susceptible cultivar Cocodrie. The association of host plant characteristics to oviposition preference is discussed. Projected oviposition patterns relative to sugarcane and rice production areas were estimated for Texas and Louisiana based on the availability of each host in different regions of each state. These results suggest that, where sugarcane and rice co-occur, the majority of eggs would be found on sugarcane early in the season, because of this crop's substantially greater biomass compared with rice. Abundance later in the season would also favor sugarcane; however, the abundance on rice would be greater than expected solely based on host availability, largely because of the greater preference per gram of rice plant dry weight.     

Second generation ethanol in Brazil: can it compete with electricity production?

Much of the controversy surrounding second generation ethanol production arises from the assumed competition with first generation ethanol production; however, in Brazil, where bioethanol is produced from sugarcane, sugarcane bagasse and trash will be used as feedstock for second generation ethanol production. Thus, second generation ethanol production may be primarily in competition with electricity production from the lignocellulosic fraction of sugarcane. A preliminary technical and economic analysis of the integrated production of first and second generation ethanol from sugarcane in Brazil is presented and different technological scenarios are evaluated. The analysis showed the importance of the integrated use of sugarcane including the biomass represented by surplus bagasse and trash that can be taken from the field. Second generation ethanol may favorably compete with bioelectricity production when sugarcane trash is used and when low cost enzyme and improved technologies become commercially available.     

Extraction and characterization of wax from sugarcane bagasse and the enzymatic hydrolysis of dewaxed sugarcane bagasse

Extraction of high-value products from agricultural wastes is an important component for sustainable bioeconomy development. In this study, wax extraction from sugarcane bagasse was performed and the beneficial effect of dewaxing pretreatment on the enzymatic hydrolysis was investigated. About 1.2% (w/w) of crude sugarcane wax was obtained from the sugarcane bagasse using the mixture of petroleum ether and ethanol (mass ratio of 1:1) as the extraction agent. Results of Fourier-transform infrared characterization and gas chromatography–mass spectrometry qualitative analysis showed that the crude sugarcane wax consisted of fatty fractions (fatty acids, fatty aldehydes, hydrocarbons, and esters) and small amount of lignin derivatives. In addition, the effect of dewaxing pretreatment on the enzymatic hydrolysis of sugarcane bagasse was also investigated. The digestibilities of cellulose and xylan in dewaxed sugarcane bagasse were 18.7 and 10.3%, respectively, compared with those of 13.1 and 8.9% obtained from native sugarcane bagasse. The dewaxed sugarcane bagasse became more accessible to enzyme due to the disruption of the outermost layer of the waxy materials.     

甘蔗遗传转化研究进展

甘蔗由于其本身的特殊性使其遗传转化较为困难,但利用直接转化法及不同的受体系统已在甘蔗不同品种上获得成功,农杆菌介导法已开始应用于甘蔗转化研究。对甘蔗遗传转化方法及其影响因素,基因工程现状进行了综述。     

低氮条件下甘蔗-大豆间作对甘蔗产量、品质及经济效益的影响

为探究甘蔗-大豆间作对甘蔗产量、品质及经济效益的影响,在施用尿素150 kg·hm^-2条件下,选择3个甘蔗品种(B8、ROC22及GT21)进行了甘蔗单作、甘蔗-大豆间作两种种植模式的试验.结果表明： 甘蔗-大豆间作对甘蔗的有效茎数、茎径以及原料蔗、蔗糖产量均有显著影响,而对原料蔗品质影响不大;与单作相比,间作大豆处理的宿根蔗茎径大小、有效茎数、蔗茎产量和糖产量分别提高5.1%～8.7%、7.9%～31.0%、9.0%～40.5%和5.6%～39.5%;每公顷原料蔗+大豆、糖+大豆可分别增收5.89～7.93万元和5.83～7.72万元;3个品种中,ROC22间作大豆的经济收益最高,而宿根蔗B8和GT21的产量均高于ROC22.表明甘蔗-大豆间作是减少氮肥施用、提高经济收入的有效栽培措施.     

The Brazilian experience of sugarcane ethanol industry

Biomass has gained prominence in the last few years as one of the most important renewable energy sources. In Brazil, a sugarcane ethanol program called ProAlcohol was designed to supply the liquid gasoline substitution and has been running for the last 30 yr. The federal government’s establishment of ProAlcohol in 1975 created the grounds for the development of a sugarcane industry that currently is one of the most efficient systems for the conversion of photosynthate into different forms of energy. Improvement of industrial processes along with strong sugarcane breeding programs brought technologies that currently support a cropland of 7 million hectares of sugarcane with an average yield of 75 tons/ha. From the beginning of ProAlcohol to the present time, ethanol yield has grown from 2,500 to around 7,000 l/ha. New technologies for energy production from crushed sugarcane stalk are currently supplying 15% of the electricity needs of the country. Projections show that sugarcane could supply over 30% of Brazil’s energy needs by 2020. In this review, we briefly describe some historic facts of the ethanol industry, the role of sugarcane breeding, and the prospects of sugarcane biotechnology     

Effects of biocides and rotation breaks on soil organisms associated with the poor early growth of sugarcane in continuous monoculture

Glasshouse and field experiments were conducted to determine the effects of biocides and rotation breaks on deleterious soil organisms associated with the poor early growth and subsequent yield decline of sugarcane grown in continuous monoculture. Fumigation of a soil that had been under sugarcane monoculture with minimal breaks for more than 30 years markedly improved the health and growth of the sugarcane sett and shoot root systems, increased the growth of the primary shoot and stimulated more and larger secondary shoots. It also reduced populations of culturable fungi in the rhizosphere of the sett roots and reduced colonization of the sett and shoot roots by lesion nematode (Pratylenchus zeae). Exposure of the developing sett root system for 14 days to mono-cultured sugarcane soil was sufficient to significantly retard subsequent plant growth. In field experiments, fungicide and nematicide (mancozeb + aldicarb), when applied together to land under sugarcane monoculture, was as effective as fumigation in improving early sugarcane growth and increasing sugarcane yields. Rotation breaks (alternate crops, sown pasture, bare fallow) that were in place for 54 months, increased sugarcane establishment and increased sugarcane yields to levels similar to that obtained following fumigation of land under sugarcane monoculture. Fumigation of land that had been under the rotation breaks gave plant growth responses that were in addition to that achieved by the breaks alone. A mancozeb + aldicarb treatment was as effective as fumigation in increasing sugarcane yields after a bare fallow break but accounted for only a portion of the fumigation response following the crop and pasture breaks. Improved plant nutrition may be a factor in the fumigation response following the crop and pasture breaks. Plant growth responses to fumigation and the manocozeb + aldicarb treatments that were manifested in final sugarcane yields (after one years growth) were evident as plant growth responses (sett root, shoot root and primary shoot dry weight) measured 54 days after planting. The experimental results support the concept that when sugarcane is grown as a monoculture, deleterious fungi and nematodes retard plant establishment and early plant growth and that this leads to reduced sugarcane yields.     

Perspective of the Sugarcane Industry in Brazil

The sugarcane industry in Brazil is experiencing a rapid shift towards creating the grounds for a green and sustainable biorefinary industry. After 30 years of ProAlcool, the federal government program that boosted Brazil’s sugarcane industry by creating a mandate to blend ethanol with gasoline, flex fuel engines now dominate Brazil’s automobile industry. Currently, bioethanol replaces around 30% of the gasoline consumed in the country and its demand is projected to more that double in the next 10 years. On another front, the sugarcane genomics program created by FAPESP in the late 1990s paved the way for the establishment of innovative biotechnology startup companies that attracted the attention of the largest agro-biotechnology sector companies of the world. Almost all of these companies now have their sugarcane research centers surrounding the city of Campinas, São Paulo. In addition, innovative synthetic biology companies are developing technologies to produce diesel, jet fuel and other high value molecules using sugarcane juice as a carbon source. The sugarcane industry also teamed with petrochemical companies and already established operating plants to produce bioplastics. Innovations have also occurred in the field of co-generation of electricity from sugarcane bagasse. Currently sugarcane supplies 4% of the electricity needs of the country. Collectively, these innovations suggest that Brazil’s sugarcane industry could supply over 30% of the country energy needs by 2021 and a significant fraction of new bioproducts produced by its nascent biorefinary plants.     

Genetically modified sugarcane for bioenergy generation

Sugarcane breeding has significantly progressed over the past 30 years, but attempts to further increase crop yield have been limited due to the complexity of the sugarcane genome. An alternative to boost the crop yield is the introduction of genes encoding desirable traits in the elite sugarcane cultivars. Genetically modified sugarcane with increased yield and pest and disease resistance has already proven its value not only by the increased sugar content but also for the improvement of the crop performance. However, transgene stability is still a challenge since transgene silencing seems to occur in a large proportion of genetically modified sugarcane plants. In addition, regulatory issues associated with the crop propagation model will also be a challenge to the commercial approval of genetically modified sugarcane.     

A job-shop scheduling approach for optimising sugarcane rail operations

The sugarcane transport system is very complex and uses a daily schedule, consisting of a set of locomotives runs, to satisfy the requirements of the mill and harvesters. The total cost of sugarcane transport operations is very high; over 35% of the total cost of sugarcane production in Australia is incurred in cane transport. Producing efficient schedules for sugarcane transport can reduce the cost and limit the negative effects that this system can have on the raw sugar production system. In this paper, the sugarcane rail operations are formulated as a blocking job shop scheduling problem. A mixed integer programming approach is used to formulate the shop job scheduling problem. Mixed integer programming and constraint programming search techniques are integrated for solving the problem. A case study is solved to test the approach.     

An-Overview on invertase in sugarcane

Saccharum officinarum is one of the most cultivated hybrid varieties among the sugarcane varieties. In sugarcane plant sucrose is the major carbohydrate which can be stored and transported. Different physiological and biochemical studies on this crop report that invertase activity and sucrose concentration some how are key limiting step in the process of sucrose accumulation. Significant efforts have been made in relation to the sucrose cycle by altering the sucrose phosphate synthetase, sucrose synthetase and invertase. In sugarcane two types of invertase enzymes have been reported on the basis of pH and cellular localization. Invertase breaks the sucrose into hexoses as a source of energy and carbon. It has also been reported that this enzyme is involved in the process of cell differentiation and plant development. Progress has been made for the understanding of invertase activity and its role in sugarcane plant. With the help of biotechnology it is possible to target the desired gene with genetic engineering approach to increase sucrose content by careful manipulation of invertase (enzyme) gene to increase the sucrose yield in sugarcane. Purpose of this mini review is to high-light the role of invertase in sugarcane and how to overcome sucrose recovery in sugarcane.     

不同土壤类型下甘蔗根系AM真菌多样性及与土壤因子关系

甘蔗是广西重要的糖料作物,本研究通过建立克隆文库、土壤养分分析和根样染色等方法测定了33个采样点3种土壤类型(赤红壤,红壤,砖红壤)下甘蔗根系AM真菌多样性及其与土壤因子的关系。结果表明,3种土壤类型的甘蔗根系共鉴定出6科6属11种AM真菌,AM真菌各属频度存在明显差异,其中球囊霉属的频度值最高,在33个根系样品中有32个存在该属,为广西甘蔗根系AM真菌的优势属,而类球囊霉属、无梗囊霉属、近明球囊霉属、多样孢囊霉属和盾巨孢囊霉属5个属为稀有属。3种土壤类型的甘蔗根系均发现有球囊霉属和盾巨孢囊霉属;近明球囊霉属、无梗囊霉属和类球囊霉属仅在赤红壤的甘蔗根系中出现;而多样孢囊霉属仅在在赤红壤和砖红壤的甘蔗根系中出现。土壤的pH与AM菌根侵染率呈显著正相关,而有机质、总N、有效P、交换性Mg2+与AM菌根侵染率均呈负相关。本研究表明,自然条件下甘蔗根系具有相对丰富的AM真菌类群,这些AM真菌可能在甘蔗生长过程中发挥着重要的生态功能。     

The Mo- and Fe-nitrogenases of the endophyte Kosakonia sp. UYSO10 are necessary for growth promotion of sugarcane

Sugarcane is a multipurpose crop primarily used to produce sugar, energy and bioethanol. It requires high amounts of N-fertilization for optimal growth, which increases production costs and environmental degradation. The contribution of biological nitrogen fixation to Uruguayan commercial sugarcane cultivars was demonstrated previously, and diazotrophic bacteria that were isolated from the stems were characterized and identified. From this collection, the isolate UYSO10 related to the Kosakonia genus (formerly Enterobacter) was described as a plant growth-promoting endophyte of sugarcane plants. To evaluate the effect of the inoculation of wild-type and nitrogenase-deficient strains of Kosakonia sp. UYSO10 on sugarcane growth promotion under non-sterile conditions. Kosakonia sp. UYSO10 was inoculated onto sugarcane setts for plant growth promotion greenhouse experiments. Single and double mutants resulting to the nitrogenase-encoding genes (nifH, anfH) were constructed, and the phenotypes were evaluated in vitro and in vivo. Kosakonia sp. UYSO10 is able to promote sugarcane growth under non-sterile conditions, that strain UYSO10 harbors two functional nitrogenases and the inactivation of both nitrogenase-encoding genes diminish its capacity of promoting growth on sugarcane. All together, the results obtained showed that the biological nitrogen fixation ability of Kosakonia sp. UYSO10 is required for sugarcane growth promotion.     

甘蔗/大豆间作减量施氮对甘蔗产量、品质及经济效益的影响

在大田栽培条件下,研究了减量施氮对广州地区不同甘蔗/大豆间作模式下甘蔗产量、品质、生物量动态变化及甘蔗群体经济效益的影响.结果表明:氮肥和种植模式对甘蔗产量、品质均无显著影响,各处理甘蔗产量、品质均无显著差异.不同施氮水平的甘蔗/大豆间作系统较单作甘蔗提高了土地利用率,其土地当量比(LER)为1.36～2.12.各处理甘蔗总生物量的动态积累随生育期的推进均符合"S"型生长曲线,低氮水平下问作甘蔗的生物量动态积累特征参数最协调,高氮水平下,甘蔗生长高峰期提前,快速生长最大持续期缩短,甘蔗产量下降,因此,可以通过调整氮素的投入量来改善生物量的增长参数,从而获得高产.低氮处理甘蔗/大豆间作系统经济效益比高氮处理高3.2%～26.3%,说明减量施氮可有效提高群体经济效益.甘蔗/大豆1∶2间作模式的经济效益最好.     

Sugarcane improvement: how far can we go?

In recent years, efforts to improve sugarcane have focused on the development of biotechnology for this crop. It has become clear that sugarcane lacks tools for the biotechnological route of improvement and that the initial efforts in sequencing ESTs had limited impact for breeding. Until recently, the models used by breeders in statistical genetics approaches have been developed for diploid organisms, which are not ideal for a polyploid genome such as that of sugarcane. Breeding programs are dealing with decreasing yield gains. The contribution of multiple alleles to complex traits such as yield is a basic question underlining the breeding efforts that could only be addressed by the development of specific tools for this grass. However, functional genomics has progressed and gene expression profiling is leading to the definition of gene networks. The sequencing of the sugarcane genome, which is underway, will greatly contribute to numerous aspects of research on grasses. We expect that both the transgenic and the marker-assisted route for sugarcane improvement will contribute to increased sugar, stress tolerance, and higher yield and that the industry for years to come will be able to rely on sugarcane as the most productive energy crop.     

甘蔗茎杆特异表达基因启动子的克隆及初步分析

甘蔗茎秆是利用转基因方法生产重组药用蛋白或有价值的化合物的理想器官,构建能在甘蔗茎秆中高水平表达异源蛋白质的表达载体是非常有意义的。而一个高效表达的载体,启动子则是其最重要元件之一,因此,茎秆特异性启动子的获得是甘蔗作为生物反应器的前提。利用染色体步移法克隆到甘蔗己糖转运蛋白基因PST2a 5′端上游的一段长1968bp的序列（ Ppst2a ）,经序列测定及软件分析表明,该序列具有典型的启动子结构。此序列置换植物表达载体pCAMBIA1301上的CaMV 35S启动子,构建植物表达载体,命名为pCAMBIA1900,该启动子下游为gus基因。利用基因枪法转化甘蔗的茎和叶,对gus基因的瞬时表达进行测定,结果表明所获得的己糖转运蛋白基因启动子只在甘蔗茎中驱动gus基因瞬时表达,该启动子具有茎秆特异性。