甘蔗幼叶脱分化及愈伤组织形成过程中内源细胞分裂素和ABA的变化

甘蔗幼叶片,叶鞘和幼茎在含有2.4-D的培养条件下脱分化的情况有明显差异。它们除了在形态学上有区别外,这三种材料原有的细胞分裂素和ABA的水平明显不同。在幼叶片脱分化及愈伤组织形成过程中,内源细胞分裂素和ABA的水平也发生了明显变化。据此我们认为在甘蔗组织培养中2.4-D可能通过调节内源激素的水平及其相互作用,引起培养物中某些生理生化过程发生改变,从而进行脱分化和愈伤组织形成。     

Biodegradation and detoxification of dyes and industrial effluents by Ganoderma weberianum B-18 immobilized in a lab-scale packed-bed bioreactor

White-rot fungi are considered to be promising biotechnological tools to complement or replace the current technologies for the treatment of effluents from textile production plants. The aim of this work was to investigate the decolorization capacity of Ganoderma weberianum B-18 in solid state fermentation with sugarcane bagasse as a substrate and ligninolytic inducer as well as to decolorize and detoxify industrial effluents by this strain in a laboratory scale packed-bed bio-reactor. The results demonstrated that G. weberianum B-18 indeed showed to possess decolorization capacity in solid state fermentation with sugarcane bagasse supplemented with synthetic dyes. Moreover, fungal biomass of G. weberianum B-18 immobilized in sugarcane bagasse in a packed-bed bioreactor was shown to efficiently decolorize and detoxify different dyes and authentic industrial effluents in semi-continuous conditions. In this decolorization process, laccase enzymes secreted by the fungus played the main role. Hence, a packed-bed reactor with G. weberianum B-18 immobilized in sugarcane bagasse seems to be a suitable system for the further development of an efficient bioprocess for large-scale treatment of dye-containing wastewaters.     

Biorefinery for combined production of jet fuel and ethanol from lipid‐producing sugarcane: a techno‐economic evaluation

Replacing fossil fuels with an economically viable green alternative at scale has proved most challenging in the aviation sector. Recently sugarcane, the most productive crop on the planet, has been engineered to accumulate lipids. This opens the way for production of far more industrial vegetable oil per acre than previously possible. This study performs techno‐economic feasibility analysis of jet fuel production from this new cost efficient and high yield feedstock. A comprehensive process model for biorefinery producing hydrotreated jet fuel (from lipids) and ethanol (from sugars), with 1 600 000 MT yr^?1 lipid‐cane processing capacity, was developed in SuperPro Designer. Considering lipid‐cane development is continuing for higher oil concentrations, analysis was performed with lipid‐cane containing 5%, 10%, 15%, and 20% lipids. Capital investments for the biorefinery ranged from 238.1 to 351.2 million USD, with jet fuel capacities of 12.6–50.5 million liters (correspondingly ethanol production of nil to 102.6 million liters). The production cost of jet fuel for different scenarios was estimated Replacing fossil fuels with an economically viable green alternative at scale has proved most challenging in the aviation sector. Recently sugarcane, the most productive crop on the planet, has been engineered to accumulate lipids. This opens the way for production of far more industrial vegetable oil per acre than previously possible. This study performs techno‐economic feasibility analysis of jet fuel production from this new cost efficient and high yield feedstock. A comprehensive process model for biorefinery producing hydrotreated jet fuel (from lipids) and ethanol (from sugars), with 1 600 000 MT yr^?1 lipid‐cane processing capacity, was developed in SuperPro Designer. Considering lipid‐cane development is continuing for higher oil concentrations, analysis was performed with lipid‐cane containing 5%, 10%, 15%, and 20% lipids. Capital investments for the biorefinery ranged from 238.1 to 351.2 million USD, with jet fuel capacities of 12.6–50.5 million liters (correspondingly ethanol production of nil to 102.6 million liters). The production cost of jet fuel for different scenarios was estimated $0.73 to $1.79 per liter ($2.74 to $6.76 per gal) of jet fuel. In all cases, the cost of raw materials accounted for more than 70% of total operational cost. Biorefinery was observed self‐sustainable for steam and electricity requirement, because of in‐house steam and electricity generation from burning of bagasse. Minimum fuel selling prices with a 10% discount rate for 20% lipid case was estimated $1.40/L ($5.31/gal), which was lower than most of the reported prices of renewable jet fuel produced from other oil crops and algae. Along with lower production costs, lipid‐cane could produce as high as 16 times the jet fuel (6307 L ha^?1) per unit land than that of other oil crops and do so using low‐value land unsuited to most other crops, while being highly water and nitrogen use efficient.     

Saccharum spontaneum L. ‘SES 208’ genetic linkage map combining RFLP- and PCR-based markers

A 527 marker linkage map ofSaccharum spontaneum L. SES 208 (2n = 64) was established by analyzing 208 single-dose (SD) arbitrarily primed PCR polymorphisms, 234 SD RFLPs, 41 double-dose (DD) and one triple-dose (TD) polymorphisms. A map hypothesis constructed using these markers (minimum LOD = 4.00, = 0.25 M) had 64 linkage groups with 13 SD, nine DD, and one TD markers unlinked. Eight chromosome homology groups were identified by using DD fragments as well as SD RFLPs that identified more than one linkage group. Linkages in repulsion phase were absent from the map, as found in two previous genetic studies of this species. Together, these data demonstrate that SES 208 displayed polysomic segregation, a genetic behavior typical of autopolyploid species. As with previous studies, it was concluded that SES 208 behaved like an auto-octoploid, which was also in agreement with the number of homology groups observed. A ² was used to test whether the 527 markers were randomly distributed throughout the genome: both arbitrarily primed PCR markers and RFLPs had a distribution that was statistically indistinguishable from random. The integrated arbitrarily primed PCR-RFLP map had a predicted genomic coverage of 93% (considering only 442 SD polymorphisms) and an average interval between markers of 6 cM. SD markers were used to estimate the genome size of SES 208 at ca. 33 00 cM.     

Immobilized cyanobacteria as a biofertilizer for rice crops; Intl. Conference on Applied Algology, Knysna, South Africa, April 1996.

N₂-fixing cyanobacteria (Anabaena azollae, symbiont strains) were immobilized in polyurethane foam and ammonia production by the cyanobacteria was investigated in the laboratory and rice field. The cyanobacterial symbiont, A. azollae - MPK-SK-AM-24 showed the highest growth rate and biomass production amongst the 5 isolates examined while A. azollae-AS-DS showed the highest nitrogenase activity followed by A. variabilis - SA₀ (wild type, non-symbiotic). Treatment of the foam-immobilized cyanobacteria with the systemic fungicide Bavistin stimulated nitrogenase activity while inhibiting glutamine synthetase (GS) activity. Free-living A. azollae-MPK-SK-AF-38, A. azollae - MPK-SK-AM-24 and A. azollae-MPK-SK-AM-27 excreted the highest amounts of ammonia into the growth medium; under foam - immobilized conditions the ammonia production increased further. Treatment of the foam - immobilized cyanobacteria with the fungicides Bavistin and Vitavax resulted in ammonia production at significantly higher rates. Rice seedlings (var. ADT 36) grown in the laboratory in conjunction with foam - immobilized A. azollae showed increased growth. A field experiment with paddy rice and foam - immobilized A. azollae strains indicated that the cyanobacteria excreted significant amounts of ammonia into the flood water in the rice fields resulting in increased chlorophyll content of the plants and increased the rice grain and straw yields. A combination of fertilizer nitrogen and inoculation with foam - immobilized cyanobacteria also significantly increased the rice grain and straw yield. Additionally, both A. azollae and A. variabilis were immobilized in sugarcane waste (bagasse), added to rice paddy and resulted in increased rice grain yield. This revised version was published online in September 2006 with corrections to the Cover Date.     

Vacuolar targeting of r‐proteins in sugarcane leads to higher levels of purifiable commercially equivalent recombinant proteins in cane juice

Sugarcane is an ideal candidate for biofarming applications because of its large biomass, rapid growth rate, efficient carbon fixation pathway and a well‐developed storage tissue system. Vacuoles occupy a large proportion of the storage parenchyma cells in the sugarcane stem, and the stored products can be harvested as juice by crushing the cane. Hence, for the production of any high‐value protein, it could be targeted to the lytic vacuoles so as to extract and purify the protein of interest from the juice. There is no consensus vacuolar‐targeting sequence so far to target any heterologous proteins to sugarcane vacuole. Hence, in this study, we identified an N‐terminal 78‐bp‐long putative vacuolar‐targeting sequence from the N‐terminal domain of unknown function (DUF) in Triticum aestivum 6‐SFT (sucrose: fructan 6‐fructosyl transferase). In this study, we have generated sugarcane transgenics with gene coding for the green fluorescent protein (GFP) fused with the vacuolar‐targeting determinants at the N‐terminal driven by a strong constitutive promoter (Port ubi882) and demonstrated the targeting of GFP to the vacuoles. In addition, we have also generated transgenics with His‐tagged β‐glucuronidase (GUS) and aprotinin targeted to the lytic vacuole, and these two proteins were isolated and purified from the transgenic sugarcane and compared with commercially available protein samples. Our studies have demonstrated that the novel vacuolar‐targeting determinant could localize recombinant proteins (r‐proteins) to the vacuole in high concentrations and such targeted r‐proteins can be purified from the juice with a few simple steps.     

Parameters of fermentation and rumen microbiota of Nellore steers fed with different proportions of concentrate in fresh sugarcane containing diets

The objective of this study was to evaluate the effect of a fresh sugarcane-based diet and different roughage-to-concentrate ratios (70:30, 60:40, 40:60 and 20:80) on the rumen microbiota associated with rumen fermentation parameters and the intake and apparent digestibility of nutrients in Nellore steers. Eight rumen-cannulated Nellore steers (331 ± 8 kg BW) were distributed in a double 4 × 4 Latin square design balanced for the control of the residual effect. The ruminal pH decreased (p < 0.01) and the concentrations of N–NH₃, isovaleric and valeric acids increased linearly (p < 0.05) with an increase dietary concentrate level. Furthermore, an increased concentrate proportion reduced the population of Fibrobacter succinogenes and Ruminococus flavefaciens (p < 0.01) and increased the population of Selenomonas ruminantium and Megasphaera elsdenii (p < 0.01). The protozoa count revealed a predominance of the genus Entodinium. The synthesis of microbial N [g/d] and the efficiency of microbial synthesis [g of microbial N/kg of organic matter apparently digested in the rumen] increased as the proportion of concentrate was increased (p < 0.05). Therefore, it can be concluded that an increasing proportion of concentrate in sugarcane-containing diets enhances the synthesis of microbial protein and does not alter the fibre digestibility, although the population of fibre fermenting bacteria was reduced.     

Value of cane trash in nitrogen nutrition of sugarcane

The significance of trash containing 0.3 to 0.5% N in the N nutrition of sugarcane (Saccharum hybrid sp.) was investigated in pot- and field experiments using¹⁵N-labelled trash. The data obtained from the pot study with 2 silty-clay loams (a Humic Nitosol and a Humic Acrisol) showed that surface-applied trash (10 tonnes/ha), although ground to pass a 1-mm sieve, contributed less than 10% of N removed by sugarcane. Uptake of trash N was most active during the initial 6 months of the experiment though at the end of the study period of 18 months less than 15% of trash N was altogether recovered by sugarcane. In the absence of fertilizer N in a field study on the Humic Acrisol (C/N ratio 22), unground trash (5 tonnes. ha⁻¹) depressed soil N uptake by sugarcane by immobilizing available soil N. The field study moreover confirmed that the contribution of trash N in the supply of N to sugarcane is negligible. The value of trash would reside in its capacity to increase over the long term the organic matter level in the soils.