Factors modulating the levels of the allelochemical sorgoleone in Sorghum bicolor

Sorgoleone is the major component of the hydrophobic root exudate of sorghum [Sorghum bicolor (L.) Moench]. The presence of this allelochemical is intrinsically linked to root growth and the development of mature root hairs. However, factors modulating root formation and the biosynthesis of sorgoleone are not well known. Sorgoleone production was independent of early stages of plant development. The optimum temperature for root growth and sorgoleone production was 30°C. Seedling development and sorgoleone levels were greatly reduced at temperatures below 25°C and above 35°C. The level of sorgoleone was also sensitive to light, being reduced by nearly 50% upon exposure to blue light (470 nm) and by 23% with red light (670 nm). Applying mechanical pressure over developing seedlings stimulated root formation but did not affect the biosynthesis of this lipid benzoquinone. Sorgoleone production did not change in seedlings exposed to plant defense elicitors. On the other hand, sorgoleone levels increased in plants treated with a crude extract of velvetleaf (Abutilon theophrasti Medik.) root. This stimulation was not associated with increased osmotic stress, since decreases in water potential (Ψ_w) by increasing solute concentrations with sorbitol reduces sorgoleone production. Sorgoleone production appears to be constitutively expressed in young developing sorghum plants. Other than with temperature, changes in the environmental factors had either no effect or caused a reduction in sorgoleone levels. However, the stimulation observed with velvetleaf root crude extract suggests that sorghum seedlings may respond to the presence of other plants by releasing more of this allelochemical.     

Biosynthesis of lipid resorcinols and benzoquinones in isolated secretory plant root hairs

The primary functions of root hairs are to increase the root surface area and to aid plants in water and nutrient uptake. However, some root hairs also have secretory functions and exude bioactive secondary metabolites. Sorghum (Sorghum bicolor) root hairs release a substantial amount of phenolic lipids including sorgoleone, a 3-pentadecatriene benzoquinone. The activity of the key enzymes involved in the biosynthesis of lipid resorcinols and benzoquinones was measured directly in isolated root hair preparations obtained from 6-d-old roots. The purified root hair preparation readily converted long-chain acyl-CoA starter units to their corresponding lipid resorcinols and decanoyl-CoA was the best substrate, yielding a 5-n-nonyl-resorcinol. The isolated root hair preparation also had high S-adenosyl-L-methionine-dependent O-methyltransferase activity, which catalyses the methylation of several 5-n-alkyl-resorcinols. Optimum activity was with 5-n-pentyl-resorcinol. Isolated root hairs also exhibited hydroxylase activity (putatively a P450 mono-oxygenase) that reacted with the lipophilic 5-pentadecyl-resorcinol substrate. The in situ hydroxylase activity was low relative to the other enzymes studied, but was still detectable in isolated root hairs. Thus, sorghum root hairs possess the entire metabolic machinery necessary for the biosynthesis of lipid resorcinols and benzoquinones. This will have implications for the genetic engineering of bioactive lipid resorcinols and benzoquinones in sorghum and in other plant species. It also demonstrates that some root hairs can function as specialized cells for the production of bioactive secondary metabolites.     

Sorgoleone

Sorgoleone, a major component of the hydrophobic root exudate of sorghum [Sorghum bicolor (L.) Moench], is one of the most studied allelochemicals. The exudate also contains an equivalent amount of a lipid resorcinol analog as well as a number of minor sorgoleone congeners. Synthesis of sorgoleone is constitutive and compartmentalized within root hairs, which can accumulate up to 20 μg of exudate/mg root dry weight. The biosynthesis pathway involves unique fatty acid desaturases which produce an atypical 16:3 fatty acyl-CoA starter unit for an alkylresorcinol synthase that catalyzes the formation of a pentadecatrienylresorcinol intermediate. This intermediate is then methylated by SAM-dependent O-methyltransferases and dihydroxylated by cytochrome P450 monooxygenases. An EST data set derived from a S. bicolor root hair-specific cDNA library contained all the candidate sequences potentially encoding enzymes involved in the sorgoleone biosynthetic pathway. Sorgoleone interferes with several molecular target sites, including inhibition of photosynthesis in germinating seedlings. Sorgoleone is not translocated acropetally in older plants, but can be absorbed through the hypocotyl and cotyledonary tissues. Therefore, the mode of action of sorgoleone may be the result of inhibition of photosynthesis in young seedlings in concert with inhibition of its other molecular target sites in older plants. Due to its hydrophobic nature, sorgoleone is strongly sorbed in soil which increases its persistence, but experiments show that it is mineralized by microorganisms over time.     

WATER RELATIONS OF WATER-STRESSED,SPLIT-ROOT C4 (SORGHUM BICOLOR; POACEAE) AND C3 (HELIANTHUS ANNUUS; ASTERACEAE) PLANTS

Sorghum [Sorghum bicolor (L.) Moench] and sunflower (Helianthus annuus L.) were grown in a greenhouse with roots divided between sand irrigated with nutrient solution (–0.097 MPa) or nutrient solution containing polyethylene glycol (PEG) (–0.570 MPa) to compare the effect of unequal root zone stress on plant water relations of a C₄ (sorghum) and a C₃ (sunflower) plant. Roots also were divided between two pots of sand irrigated only with nutrient solution (controls) or only with PEG in nutrient solution. In addition to plant water-status measurements, photosynthetic rate, growth (height, root, and shoot dry weights), and evolution of ethylene (a gaseous hormone indicative of stress) were measured. Under all three split-root treatments, sunflower had a lower leaf water potential and produced more ethylene than sorghum. Sunflower was able to survive the PEG stress if half of its root system was under nonstressed conditions. Sunflower with half its root system irrigated with PEG usually had values of leaf water potential, osmotic potential, stomatal resistance, transpiration rate, photosynthetic rate, ethylene evolution, height, and dry weights that were close to those of the control plants. Sunflower with all roots exposed to PEG was wilted severely. Sorghum was little affected by PEG stress applied either to half or all the root system. Growth of sorghum was the same under all treatments. Apparently because stomata of sorghum were more closed in the partial stress test than those of sunflower, sorghum conserved water and had a higher leaf water potential, which might have permitted growth with stress.     

In planta transformation of sorghum (Sorghum bicolor (L.) Moench) using TPS1 gene for enhancing tolerance to abiotic stresses

Journal of Genetics - An in planta transformation protocol for sorghum (Sorghum bicolor (L.) Moench) using shoot apical meristem of germinating seedlings is reported in this study. Agrobacterium...     

Molecular genetic analysis of soriz genome (Sorghum oryzoidum)

Molecular-genetic analysis of soriz genotypes (Sorghum oryzoidum), its paternal form Sorghum bicolor (L.) Moench (grain sorghum), possible parents (Sorghum sudanense (Piper.) Stapf. (Sudan grass) and Oryza sativa L. (rice planting)) and the nearest relatives has been carried out using microsatellite (MS) loci of sorghum and rice. Based on these data genetic distances have been calculated. It was shown that soriz do not bear DNA fragments of rice, but contains in its genome DNA fragments belonging to the Sudanese grass indicating that the origin of soriz is associated with Sorghum sudanense.     

Functional characterization of desaturases involved in the formation of the terminal double bond of an unusual 16:3Delta(9,12,150) fatty acid isolated from Sorghum bicolor root hairs

Sorgoleone, produced in root hair cells of sorghum (Sorghum bicolor), is likely responsible for much of the allelopathic properties of sorghum root exudates against broadleaf and grass weeds. Previous studies suggest that the biosynthetic pathway of this compound initiates with the synthesis of an unusual 16:3 fatty acid possessing a terminal double bond. The corresponding fatty acyl-CoA serves as a starter unit for polyketide synthases, resulting in the formation of 5-pentadecatrienyl resorcinol. This resorcinolic intermediate is then methylated by an S-adenosylmethionine-dependent O-methyltransferase and subsequently dihydroxylated, yielding the reduced (hydroquinone) form of sorgoleone. To characterize the corresponding enzymes responsible for the biosynthesis of the 16:3 fatty acyl-CoA precursor, we identified and cloned three putative fatty acid desaturases, designated SbDES1, SbDES2, and SbDES3, from an expressed sequence tag (EST) data base prepared from isolated root hairs. Quantitative real-time RT-PCR analyses revealed that these three genes were preferentially expressed in sorghum root hairs where the 16:2 and 16:3 fatty acids were exclusively localized. Heterologous expression of the cDNAs in Saccharomyces cerevisiae revealed that recombinant SbDES2 converted palmitoleic acid (16:1Delta(9)) to hexadecadienoic acid (16:2Delta(9,12)), and that recombinant SbDES3 was capable of converting hexadecadienoic acid into hexadecatrienoic acid (16:3Delta(9,12,15)). Unlike other desaturases reported to date, the double bond introduced by SbDES3 occurred between carbons 15 and 16 resulting in a terminal double bond aliphatic chain. Collectively, the present results strongly suggest that these fatty acid desaturases represent key enzymes involved in the biosynthesis of the allelochemical sorgoleone.     

Nitrate Reductase Activity: A Biochemical Criterion of Hybrid Vigour in Sorghum bicolor (L.) Moench

F₁ hybrids of Sorghum bicolor (L.) Moench and their inbred parentswere analysed for NADH-nitrate reductase activity during theearly stages of seedling growth. In all the hybrids both mid-parentaland better parental heterosis were discernible in shoots whereasin roots two hybrids out of the three tested, showed heteroticlevels. It is suggested that in sorghum nitrate reductase activityduring seedling stages can be used as a biochemical criterionfor evaluating hybrid vigour. Sorghum bicolor (L.) Moench, sorghum, hybrid vigour, nitrate reductase     

Application of silicon enhanced drought tolerance in Sorghum bicolor

The effects of silicon application on the drought tolerance of sorghum ( Sorghum bicolor (L.) Moench) were investigated for two cultivars differing in drought susceptibility. Silicon application ameliorated the decrease in dry weight under drought stress conditions, but had no effect on dry matter production under wet conditions. Under dry conditions, silicon-applied sorghum had a lower shoot to root (S/R) ratio, indicating the facilitation of root growth and the maintenance of the photosynthetic rate and stomatal conductance at a higher level compared with plants grown without silicon application. The diurnal determination of the transpiration rate indicated that the silicon-applied sorghum could extract a larger amount of water from drier soil and maintain a higher stomatal conductance. Very similar effects of silicon application were observed for both cultivars regardless of their drought susceptibility. These results suggest that silicon application may be useful to improve the drought tolerance of sorghum via the enhancement of water uptake ability.     

Incorporation and translocation of 2-deoxy-2-[18F]fluoro-d-glucose in Sorghum bicolor (L.) Moench monitored using a planar positron imaging system

[¹⁸F]FDG (2-deoxy-2-[¹⁸F]fluoro-d-glucose) was fed to a sorghum plant [Sorghum bicolor (L.) Moench] from the tip of a leaf and its movement was monitored using a planar positron imaging system (PPIS). [¹⁸F]FDG was uptaken from the leaf tip and it was translocated to the basal part of the shoots from where it moved to the roots, the tillers and the sheaths. Autoradiographic analysis of the distribution of ¹⁸F, [¹⁸F]FDG and/or its metabolites showed translocation to the roots, tillers, and to the leaves that were younger than the supplied leaf. Strong labelling was observed in the basal part of the shoots, in the sheaths, the youngest leaf and the root tips. Our results indicate that [¹⁸F]FDG and/or its metabolites were absorbed from the leaf and translocated to the sites where nutrients are required. This strongly suggests that [¹⁸F]FDG can be utilised as a tracer to study photoassimilate translocation in the living plant. This is the first report on the use of [¹⁸F]FDG, which is routinely used as a probe for clinical diagnosis, to study source to sink translocation of metabolites in whole plants in real time.     

Preservation of potential fermentables in sweet sorghum by ensiling

Pressed and wilted samples of sweet sorghum [Sorghum bicolor (L.) Moench var. Rio] were ensiled for periods up to 155 days. A kinetic study of the biochemical changes which occurred during ensiling showed that in wilted sorghum ensilage invert sugars and mannitol levels collectively were maintained at 65% of the original ferment able sugar content of the sorghum. The acidic environment produced by ensiling also served as a pretreatment that resulted in enhanced yields of reducing sugar when the sorghum was contacted with cellulolytic enzymes. The quantity of sugar obtained from enzymatic hydrolysis more than compensated for carbohydrate used by organisms during the ensiling process. Both Saccharomyces uvarum and Clostridium acetobutylicum were able to ferment a medium constituted from pressed sorghum juice and the solution resulting from enzymatic hydrolysis of sweet sorghum ensilage.     

高粱SSA-1无融合生殖胚胎学研究

经常规石蜡切片法,在光学水平观察了高粱（Ｓｏｒｇｈｕｍ ｂｉｃｏｌｏｒ （Ｌ．） Ｍｏｅｎｃｈ） ＳＳＡ－１ 无融合生殖的胚胎发生。高粱ＳＳＡ－１ 的无融合生殖为无孢子生殖和二倍体孢子生殖两种类型。两种生殖类型的单核胚囊经３ 次有丝分裂形成７ 细胞（８ 核）的成熟胚囊,由卵细胞、２ 个助细胞、２ 个极核和３ 个反足细胞组成。反足细胞迅速分裂增殖,形成由２０—３０ 个细胞组成的细胞团。此外,还具有一定频率的无孢子生殖多孢原和多胚囊现象。在未授粉情况下,卵细胞自发分裂形成典型的禾本科类型单子叶胚。经切片统计表明,ＳＳＡ－１ 的无融合生殖频率为４２％ ,证明该系为一兼性无融合生殖系。文中还讨论了ＳＳＡ－１ 无融合生殖过程的特点。     

Mapping malting quality and yield characteristics in a north American two-rowed malting barley × wild barley advanced backcross population

Molecular Breeding - The brown midrib (bmr) phenotype is a recessive trait of sorghum (Sorghum bicolor L. Moench) that results in overall lignin reduction and is associated with enhanced ruminant...     

Degradation of crude oil in the rhizosphere of Sorghum bicolor

Dissipation of petroleum contaminants in the rhizosphere is likely the result of enhanced microbial degradation. Plant roots may encourage rhizosphere microbial activity through exudation of nutrients and by providing channels for increased water flow and gas diffusion. Phytoremediation of crude oil in soil was examined in this study using carefully selected plant species monitored over specific plant growth stages. Four sorghum (Sorghum bicolor L.) genotypes with differing root characteristics and levels of exudation were established in a sandy loam soil contaminated with 2700 mg crude oil/kg soil. Soils were sampled at three stages of plant growth: five leaf, flowering, and maturity. All vegetated treatments were associated with higher remediation efficiency, resulting in significantly lower total petroleum hydrocarbon concentrations than unvegetated controls. A relationship between root exudation and bioremediation efficiency was not apparent for these genotypes, although the presence of all sorghum genotypes resulted in significant removal of crude oil from the impacted soil.     

Silicon-induced changes in viscoelastic properties of sorghum root cell walls

Silicon is deposited in the endodermal tissue in sorghum (Sorghum bicolor L. Moench) roots. Its deposition is thought to protect vascular tissues in the stele against invasion by parasites and drying soil via hardening of endodermal cells. We studied the silicon-induced changes in mechanical properties of cell walls to clarify the role of silicon in sorghum root. Sorghum seedlings were grown in nutrient solution with or without silicon. The mechanical properties of cell walls were measured in three separated root zones: basal, apical and subapical. Silicon treatment decreased cell-wall extensibility in the basal zone of isolated stele tissues covered by endodermal inner tangential walls. The silicon-induced hardening of cell walls was also measured with increases in elastic moduli (E) and viscosity coefficients (eta). These results provided new evidence that silicon deposition might protect the stele as a mechanical barrier by hardening the cell walls of stele and endodermal tissues. In contrast to the basal zone, silicon treatment increased cell-wall extensibility in the apical and subapical zones with concomitant decrease in E and eta. Simultaneously, silicon promoted root elongation. When root elongation is promoted by silicon, one of the causal factors maybe the silicon-enhanced extensibility of cell walls in the growing zone.     

phyB-1 sorghum maintains responsiveness to simulated shade, irradiance and red light: far-red light

The sorghum [Sorghum bicolor (L.) Moench] phyB-1 mutant exhibits a constitutive shade-avoidance phenotype including excessive shoot elongation. It was previously shown that this mutant also overproduces ethylene. Although phytochrome B (phyB) is assumed to be the pigment most important in sensing and transducing shade signals, the sorghum phyB-1 mutant still responds to light signals characteristic of shade. Specifically, it was determined that the leaf blade : leaf sheath elongation of phyB-1 is responsive to red : far red (R : FR), but this response is opposite that of wild type (WT). Reducing the photosynthetic photon flux density (PPFD) strongly reduced the leaf blade : leaf sheath of WT but did not affect phyB-1, demonstrating a role for phyB in sensing PPFD. Using light-emitting diode (LED) lighting, it was found that WT ethylene production was increased with low R : FR while PPFD had no effect. Conversely, phyB-1 ethylene production increased only with high PPFD, high R : FR which was the treatment resulting in the least ethylene production by WT. Elevated ethylene production inhibits shoot elongation, but may contribute to shade avoidance by reducing leaf blade : leaf sheath elongation. Ethylene responses to light treatments designed to promote or reduce phytochrome A (phyA) activity, and the analysis of PHYA levels in the two cultivars suggests that phyA could be involved in transducing shade signals in light-grown sorghum. Responses potentially tranduced by phyA are elevated in phyB-1 which also over-expresses PHYA.     

Biochemical composition and photosynthetic activity of chloroplasts from Striga hermonthica and Striga aspera, root parasites of field-grown cereals

Striga hermonthica (Del.) Benth. and Striga aspera (Willd.) Benth. are root parasites causing dramatic losses in field-grown cereals in semi-and tropics. Being achlorophyllous and obligate parasites during their underground development, upon emergence from the soil, they become green leafy plants; but, despite the presence of chlorophyll, they exhibit only low rates of photosynthesis. To investigate if deficiency in the photosynthetic apparatus could account for the low rates of photosynthesis, chloroplasts were isolated from S. hermonthica parasitizing sorghum [Sorghum bicolor (L.) Moench cv. Tiemarifing] and from S. aspera parasitizing maize ( Zea mays L. cv. Tiémantié ) grown under greenhouse conditions or in their natural surroundings. Isolated chloroplasts exhibited the characteristics of chloroplasts from C₃ plants but displayed low levels of chlorophyll and polar lipids, while the protein content was less reduced. Main changes occurred in polar lipid composition, with decreases in monogalactosyldiacylglycerol and digalactosyldiacylglycerol. All polar lipids showed a decrease in the degree of unsaturation of fatly acids. All these changes were particularly pronounced in chloroplasts from plants that experienced heavy drought in Africa. On a chlorophyll basis, chloroplasts did not display a dramatic decrease in photosynthetic activities. These results are discussed in relation to parasitism and drought adaptation.