盐地碱蓬二型性种子及其幼苗对盐渍环境的适应性

研究了盐地碱蓬二型性种子中离子含量与刚萌发幼苗耐盐性之间的关系,以及盐分对砂培盐地碱蓬二型性种子的幼苗生长、离子含量及光合特性的影响.棕色种子中离子含量显著高于黑色种子.与对照相比,100和400 mmol/L NaCl对棕色种子幼苗伸长没有抑制作用,却显著抑制黑色种子幼苗的伸长.NaCl处理下棕色种子的幼苗地上部分干重和主茎一级分枝数比黑色种子幼苗高,但二型性种子的幼苗叶片中离子含量、叶绿素含量及光合放氧速率却没有明显差异.上述结果说明盐地碱蓬棕色种子较高的离子含量可能是棕色种子刚萌发幼苗耐盐性较强的重要原因.棕色种子幼苗较高的生物量可能与其较多的分枝数有关.二型种子的这些特征可能决定了其在群落建成中所起到的不同作用.     

The effect of plant growth regulators, nitric oxide, nitrate, nitrite and light on the germination of dimorphic seeds of Suaeda salsa under saline conditions

Suaeda salsa, a leaf succulent shrub in the family Chenopodiaceae, is one of the most important halophytes in China. Suaeda salsa produces dimorphic seeds (soft brown seeds and hard black seeds). Seeds of S. salsa were collected from the coastal salt flats near Huanghua City, China. Experiments were conducted to determine the salinity-alleviating effect of plant growth regulators, nitric oxide, nitrate, nitrite and light on the germination of dimorphic seeds of S. salsa. Brown seeds had a higher germination rate than black seeds in all experiments. Black seeds were more sensitive to salt in the absence of light in comparison to brown seeds. Brown seeds absorbed water more quickly in comparison to black seeds and were found to be more tolerant of salt stress. Our results showed that 1-aminocyclopropane-1-carboxylate (ACC, the immediate precursor of ethylene), nitrite, GA₄ and BA improved seed germination in the presence of salt. However, nitrate, GA₁, GA₃ failed to alleviate salt stress. ABA inhibited seed germination and seedling growth. Possible mechanisms involved in the alleviation of salt stress in S. salsa seeds and the ecological adaptation of the seeds to the environment are discussed.     

The role of the seed coat in adaptation of dimorphic seeds of the euhalophyte Suaeda salsa to salinity

The role of the seed coat in adaptation of dimorphic seeds of the euhalophyte Suaeda salsa to salinity was investigated during germination and early seedling growth. Black and brown seeds were treated with chloroform for 1 min before the extract was used to analyze waxes and the seeds to investigate the protective role of the seed coat under saline conditions. Waxes in black seed coats were more abundant than those in brown seed coats. Salinity (500 mM NaCl) increased the concentration of Na⁺ and decreased the concentration of K⁺ in both black and brown seeds regardless of chloroform treatment. Chloroform treatment alone (in the absence of NaCl) had no effect on the concentration of Na⁺ or K⁺ in black or brown seeds and in the presence of 500 mM NaCl had no effect on the concentration of Na⁺ or K⁺ in brown seeds. However, chloroform treatment increased Na⁺ and decreased K⁺ in black seeds with 500 mM NaCl. A change of MDA (malondialdehyde) concentration in black and brown seeds treated with or without chloroform was similar to the change of Na⁺ concentration. High salinity (1500 mM NaCl) pretreatment for 40 days had a less adverse effect on germination of black seeds compared with brown seeds after they were transferred to fresh water regardless of chloroform treatment. Similar results were found for seedling emergence. In conclusion, a black seed coat may be more protective than a brown seed coat, probably by shielding the embryo from ion toxicity, because of its higher content of waxes. Thus black seeds can better maintain seed viability than brown seeds for extended periods under hypersaline conditions.     

Increase in unsaturated fatty acids in membrane lipids of Suaeda salsa L. enhances protection of photosystem II under high salinity

In order to examine the possible role of unsaturated fatty acids in photosynthesis of halophytes under high salinity, the effect of salinity on plant growth, chlorophyll (Chl) content, photochemical efficiency of PSII, membrane lipid content and fatty acids composition of a C₃ euhalophyte Suaeda salsa L. was investigated. Salt stress induced a slight increase of the maximal photochemical efficiency of PSII (F_v/F_m), actual PSII efficiency (Φ_PSII), Chl a content and Chl a/b ratio. The unsaturated fatty acid content also increased under salt stress. The proportion of MGDG, DGDG, SQDG, and PC decreased, while the proportion of PG increased from 10.9% to 26.9% under salt stress. These results suggest that S. salsa displays high resistance to photoinhibition under salt stress and that increased concentration of unsaturated fatty acids in membrane lipids of S. salsa enhances the tolerance of photosystem II to salt stress.     

Using euhalophytes to understand salt tolerance and to develop saline agriculture: Suaeda salsa as a promising model

Background As important components in saline agriculture, halophytes can help to provide food for a growing world population. In addition to being potential crops in their own right, halophytes are also potential sources of salt-resistance genes that might help plant breeders and molecular biologists increase the salt tolerance of conventional crop plants. One especially promising halophyte is Suaeda salsa, a euhalophytic herb that occurs both on inland saline soils and in the intertidal zone. The species produces dimorphic seeds: black seeds are sensitive to salinity and remain dormant in light under high salt concentrations, while brown seeds can germinate under high salinity (e.g. 600 mm NaCl) regardless of light. Consequently, the species is useful for studying the mechanisms by which dimorphic seeds are adapted to saline environments. S. salsa has succulent leaves and is highly salt tolerant (e.g. its optimal NaCl concentration for growth is 200 mm). A series of S. salsa genes related to salt tolerance have been cloned and their functions tested: these include SsNHX1, SsHKT1, SsAPX, SsCAT1, SsP5CS and SsBADH. The species is economically important because its fresh branches have high value as a vegetable, and its seed oil is edible and rich in unsaturated fatty acids. Because it can remove salts and heavy metals from saline soils, S. salsa can also be used in the restoration of salinized or contaminated saline land.Scope Because of its economic and ecological value in saline agriculture, S. salsa is one of the most important halophytes in China. In this review, the value of S. salsa as a source of food, medicine and forage is discussed. Its uses in the restoration of salinized or contaminated land and as a source of salt-resistance genes are also considered.     

Identification and evaluation of ω-3 fatty acid desaturase genes for hyperfortifying α-linolenic acid in transgenic rice seed

α-Linolenic acid (ALA) deficiency and a skewed of ω6:ω3 fatty acid ratio in the diet are a major explanation for the prevalence of cardiovascular diseases and inflammatory/autoimmune diseases. There is a need to enhance the ALA content and to reduce the ratio of linoleic acid (LA) to ALA. Six ω-3 (Δ-15) fatty acid desaturase (FAD) genes were cloned from rice and soybean. The subcellular localizations of the proteins were identified. The FAD genes were introduced into rice under the control of an endosperm-specific promoter, GluC, or a Ubi-1 promoter to evaluate their potential in increasing the ALA content in seeds. The ALA contents in the seeds of endoplasmic reticulum (ER)-localized GmFAD3-1 and OsFAD3 overexpression lines increased from 0.36 mg g?1 to 8.57 mg g?1 and 10.06 mg g?1, respectively, which was 23.8- and 27.9-fold higher than that of non-transformants. The trait of high ALA content was stably inheritable over three generations. Homologous OsFAD3 is more active than GmFAD3-1 in catalysing LA conversion to ALA in rice seeds. Overexpression of ER-localized GmFAD3-2/3 and chloroplast-localized OsFAD7/8 had less effect on increasing the ALA content in rice seeds. The GluC promoter is advantageous compared with Ubi-1 in this experimental system. The enhanced ALA was preferentially located at the sn-2 position in triacylglycerols. A meal-size portion of high ALA rice would meet >80% of the daily adult ALA requirement. The ALA-rich rice could be expected to ameliorate much of the global dietary ALA deficiency.     

Effect of salinity on germination,seedling emergence,seedling growth and ion accumulation of a euhalophyte Suaeda salsa in an intertidal zone and on saline inland

The effect of salinity on germination, seedling emergence, seedling growth and ion accumulation of a euhalophyte Suaeda salsa L. in an intertidal zone and on saline inland were investigated. Brown seeds of S. salsa were heavier and better developed than black seeds in both the intertidal zone and on saline inland. The brown seeds/black seeds ratio for S. salsa in the intertidal zone was much higher than that for S. salsa on saline inland. More germinated seeds grew as seedlings under high salinity for S. salsa from the intertidal zone than S. salsa on saline inland; high salinity decreased the shoot length more severely for S. salsa from saline inland than for S. salsa from the intertidal zone; the seedling growth at a range of NaCl, measured either as shoot length or shoot dry weight, for S. salsa from the intertidal zone was lower than that of S. salsa from saline inland. In conclusion, for S. salsa from the intertidal zone there appears to be selection for slower growth and producing more brown seeds. The establishment of populations of S. salsa in different saline environments depends on the responses of seed germination, seedling emergence and seedling growth to salinity. These characteristics may determine the natural distributions of S. salsa populations in different saline environments.     

Effects of salinity and nitrate on production and germination of dimorphic seeds applied both through the mother plant and exogenously during germination in Suaeda salsa

Salinity and nitrogen are two important environmental factors that affect the distribution of halophytes in their natural saline habitats. Seeds of the euhalophyte Suaeda salsa L. were harvested from plants that had been treated with 1 or 500 mm NaCl combined with 0.5 or 5 mm NO₃^?‐N (nitrate) for 115 days in a glasshouse. Germination was evaluated under different concentrations of NaCl and nitrate. Plants exposed to high salinity (500 mm ) and low nitrate (0.5 mm ) tended to produce heavy seeds. Either high salinity (500 mm ) or high nitrate (5 mm ) increased the brown/black seed ratio. The concentrations of Na⁺, K⁺, and Cl^? were higher in brown than in black seeds, and NO₃^? concentrations were higher in black than in brown seeds, regardless of NaCl and nitrate treatments during plant culture. Regardless of NaCl and nitrate concentrations during germination, seeds from plants grown with 0.5 mm nitrate generally germinated more rapidly than seeds from plants grown with 5 mm nitrate, and the difference was greater for black than for brown seeds. Exogenous nitrate during germination enhanced the germination of brown seeds less than that of black seeds. Producing more brown seeds and heavy black or brown seeds appears to be an adaptation of S. suaeda to saline environments. Producing more black seeds, which tend to remain dormant, should reduce competition for nitrogen and appears to be an adaptation to nitrogen‐limited environments. In conclusion, nitrate provided exogenously or by mother plants to black seeds may act as a signal molecule that enhances the germination of black S. suaeda seeds.     

温度和紫外照射对紫苏FAD7基因表达的影响

植物ω-3脂肪酸脱氢酶(ω-3FAD)基因是催化亚油酸转化为α-亚麻酸(ALA)的关键酶基因,通过调节该基因的表达,可以提高植物ALA的含量。为了研究温度和紫外照射对紫苏PfFAD7的影响,通过RTPCR方法分析了紫苏地上组织的特异性表达和温度、紫外胁迫下紫苏叶片和茎中PfFAD7基因的积累情况。分析表明,PfFAD7基因在紫苏全植株中均有表达,但在叶片中表达量最高,温度和紫外照射均影响PfFAD7基因的表达,低温可诱导PfFAD7基因的表达,而高温则抑制PfFAD7基因的表达;UV-B照射下,PfFAD7基因在叶片和茎中表达量均表现为先升高再降低。本试验对于紫苏ω-3脂肪酸脱氢酶的研究有利于高水平ALA的积累,更有利于紫苏资源的开发和利用,同时对于进一步了解不饱和脂肪酸的积累和代谢过程以及关键基因PfFAD7在此过程中的功能提供了依据。     

Increasing α-linolenic acid content in rice bran by embryo-specific expression of ω3/Δ15-desaturase gene

Rice bran, the byproduct of brown rice, with a million tons produced annually, is the major storage organ for lipids. Increasing the α-linolenic acid (ALA) content in rice bran by biotechnology strategies is beneficial for both human health and the oil industries. We introduced two ω3/Δ15 fatty acid desaturase genes cloned from rice and soybean into rice under the control of an embryo-specific promoter, REG. The ALA content (dry weight) in embryos and bran was increased up to 6.09 and 5.86 mg/g, respectively, in transgenic lines, which was 25.4- and 27.9-fold higher than the 0.24 and 0.21 mg/g in the nontransformant control. ALA accounted for 46.7 and 44.3 % of the total fatty acids in embryos and bran, respectively, of transgenic plants, which was comparable to that in linseed and perilla seeds. The trait of high ALA content was stably inherited. The enhanced ALA content was preferentially located at the sn-2 position in triacylglycerols, which are digestible and absorbable for humans. ALA-enriched rice bran may help alleviate human health problems caused by ALA deficiency, with the production of healthy bran oil for humans and feed for animals.     

Histone acetyltransferase general control non‐repressed protein 5 (GCN5) affects the fatty acid composition of Arabidopsis thaliana seeds by acetylating fatty acid desaturase3 (FAD3)

Seed oils are important natural resources used in the processing and preparation of food. Histone modifications represent key epigenetic mechanisms that regulate gene expression, plant growth and development. However, histone modification events during fatty acid (FA) biosynthesis are not well understood. Here, we demonstrate that a mutation of the histone acetyltransferase GCN5 can decrease the ratio of α‐linolenic acid (ALA) to linoleic acid (LA) in seed oil. Using RNA‐Seq and ChIP assays, we identified FAD3, LACS2, LPP3 and PLAIIIβ as the targets of GCN5. Notably, the GCN5‐dependent H3K9/14 acetylation of FAD3 determined the expression levels of FAD3 in Arabidopsis thaliana seeds, and the ratio of ALA/LA in the gcn5 mutant was rescued to the wild‐type levels through the overexpression of FAD3. The results of this study indicated that GCN5 modulated FA biosynthesis by affecting the acetylation levels of FAD3. We provide evidence that histone acetylation is involved in FA biosynthesis in Arabidopsis seeds and might contribute to the optimization of the nutritional structure of edible oils through epigenetic engineering.     

Alpha-linolenic acid and its conversion to longer chain n−3 fatty acids: Benefits for human health and a role in maintaining tissue n−3 fatty acid levels

There is little doubt regarding the essential nature of alpha-linolenic acid (ALA), yet the capacity of dietary ALA to maintain adequate tissue levels of long chain n−3 fatty acids remains quite controversial. This simple point remains highly debated despite evidence that removal of dietary ALA promotes n−3 fatty acid inadequacy, including that of docosahexaenoic acid (DHA), and that many experiments demonstrate that dietary inclusion of ALA raises n−3 tissue fatty acid content, including DHA. Herein we propose, based upon our previous work and that of others, that ALA is elongated and desaturated in a tissue-dependent manner. One important concept is to recognize that ALA, like many other fatty acids, rapidly undergoes β-oxidation and that the carbons are conserved and reused for synthesis of other products including cholesterol and fatty acids. This process and the differences between utilization of dietary DHA or liver-derived DHA as compared to ALA have led to the dogma that ALA is not a useful fatty acid for maintaining tissue long chain n−3 fatty acids, including DHA. Herein, we propose that indeed dietary ALA is a crucial dietary source of n−3 fatty acids and its dietary inclusion is critical for maintaining tissue long chain n−3 levels.     

Reproductive allocation and fruit-set pattern in the euhalophyte Suaeda salsa in controlled and field conditions

Reproductive allocation and fruit-set pattern are important characteristics of halophytes, and these characteristics were investigated in euhalophyte Suaeda salsa (S. salsa). The plants comprised of those treated with 1 mM (control) or 500 mM (high salinity) of NaCl for 120 days in a glasshouse and those collected in the field. In controlled conditions, high salinity increased reproductive allocation and the number of fruits and seeds in each node of the branches. In field conditions, reproductive allocation and the number of fruits and brown seeds in S. salsa at the upper tide line were higher than at the lower tide line. There were more fruits and seeds distributed in the central part than those in the base or top of branches in both controlled and field conditions. Soil organic matter, total nitrogen and total phosphorus content at the upper tide line were lower than at the lower tide line. An opposite trend could be observed in the total soluble salt, Na⁺ and Cl^? content in soil. In conclusion, the fruit-set pattern of S. salsa may be affected by both resource constraints and resource allocation. Reproductive allocation and fruit-set pattern may ensure the establishment of S. salsa in high saline and low nutrient environments.     

Differential salt tolerance and similar responses to nitrogen availability in plants grown from dimorphic seeds of Suaeda salsa

Suaeda salsa is an annual halophyte which produces two morphologically distinct types of seeds on the same plant. The main purpose of this study was to investigate growth responses of S. salsa plants to different levels of NaCl and nitrate nitrogen, and its significance from the viewpoint of photosynthetic physiology. In a pot experiment, we sowed seeds belonging to the two morphs into a substrate with three salinity and three nutrient levels. Plants derived from brown seeds grew well at moderate salinity (300 mmol L⁻¹ NaCl). Shoot weight of plants from black seeds gradually decreased with the increase of salinity. Plants derived from both seed morphs had the same growth rates under similar nitrogen levels. Plant growth status was generally related to chlorophyll content and photosynthetic rates. Our study shows that plants grown from the two different seed morphs of S. salsa exhibit different salt tolerance, but have similar responses to nitrate nitrogen. This is the first report on different responses to salinity and nitrogen availability in plants with heteromorphic seeds.     

FATTY ACIDS,AMINO ACIDS,MINERAL CONTENTS,AND PROXIMATE COMPOSITION OF SOME BROWN SEAWEEDS1

This study was conducted to create a nutritional database on brown seaweeds and to popularize their consumption and utilization in Iran. The fatty acid contents, amino acids profiles, and certain mineral elements composition of some brown seaweeds, Padina pavonica (L.) Thivy, Dictyota dichotoma (Huds.) J. V. Lamour., and Colpomenia sinuosa (Mert. ex Roth) Derbés et Solier were determined. Total lipid content ranged from 1.46 ± 0.38 to 2.94 ± 0.94 g · 100 g^?1dry weight (dwt), and the most abundant fatty acids were C16:0, C18:1, C20:4 ω6, and C20:5 ω3. The unsaturated fatty acids predominated in all species and had balanced sources of ω3 and ω6 acids. Highest total polyunsaturated fatty acid (PUFA) levels occurred in C. sinuosa. The protein content of D. dichotoma was 17.73 ± 0.29 g · 100 g^?1dwt, significantly higher than the other seaweeds examined. Among amino acids essential to human nutrition, methionine (Met; in D. dichotoma and P. pavonica) and lysine (Lys; in C. sinuosa) were present in high concentrations. The crude fiber content varied by 9.5 ± 11.6 g · 100 g^?1dwt in all species. Chemical analysis indicated that ash content was between 27.02 ± 0.6 and 39.28 ± 0.7 g · 100 g^?1dwt, and that these seaweeds contained higher amounts of both macrominerals (7,308–9,160 mg · 100 g^?1dwt; Na, K, Ca) and trace elements (263–1,594 mg · 100 g^?1dwt; Fe, Ni, Mn, Cu, Co) than have been reported for edible land plants. C. sinuosa had the highest amount of Ca, Fe, and a considerable content of Na was measured in P. pavonica.     

Heterologous expression of two <Emphasis Type="Italic">Glycine max</Emphasis> ω-3 fatty acid desaturases in <Emphasis Type="Italic">Saccharomyces cerevisiae</Emphasis>

α-Linolenic acid (ALA, C 18:3^Δ9,12,15) has many important biological functions. ω-3 Fatty acid desaturase (FAD3), existing in cytosolic and plastidic compartments of higher plants, catalyzes linoleic acid (LA) desaturation to produce ALA. GmFAD3A-2 and GmFAD3C genes encoding cytosolic FAD3 from Qihuang 29 soybean were cloned and inserted into p416 vector and expressed in K601 yeast strain. Gas chromatography showed that the transformed yeast strains could produce ALA. The ALA accumulation levels for the strains transformed with GmFAD3A-2 or GmFAD3C genes were 0.77 ± 0.1 and 4.13 ± 0.4% of total fatty acids, respectively, while, as compared with that of the control, the contents of LA decreased from 14.34 ± 0.8 to 10.93 ± 0.0 and 7.85 ± 0.1%, respectively, implying that the GmFAD3C enzyme is more vigorous or stable, than GmFAD3A-2. Published in Russian in Fiziologiya Rastenii, 2009, Vol. 56, No. 4, pp. 629–634. This text was submitted by the authors in English.     

Improved ethanol tolerance of Saccharomyces cerevisiae strains by increases in fatty acid unsaturation via metabolic engineering

To enhance the ethanol tolerance of Saccharomyces cerevisiae, the Arabidopsis thaliana FAD2 gene and/or the S. cerevisiae OLE1 gene were over-expressed in this yeast. The transformant over-expressing both these genes could not only synthesize dienoic fatty acids but also increased the unsaturated fatty acid content of membrane lipid and then showed the highest viability in the presence of 15% (v/v) ethanol.     

Significant enhancement of fatty acid composition in seeds of the allohexaploid,Camelina sativa,using CRISPR/Cas9 gene editing

The CRISPR/Cas9 nuclease system is a powerful and flexible tool for genome editing, and novel applications of this system are being developed rapidly. Here, we used CRISPR/Cas9 to target the FAD2 gene in Arabidopsis thaliana and in the closely related emerging oil seed plant, Camelina sativa, with the goal of improving seed oil composition. We successfully obtained Camelina seeds in which oleic acid content was increased from 16% to over 50% of the fatty acid composition. These increases were associated with significant decreases in the less desirable polyunsaturated fatty acids, linoleic acid (i.e. a decrease from ~16% to <4%) and linolenic acid (a decrease from ~35% to <10%). These changes result in oils that are superior on multiple levels: they are healthier, more oxidatively stable and better suited for production of certain commercial chemicals, including biofuels. As expected, A. thaliana T₂ and T₃ generation seeds exhibiting these types of altered fatty acid profiles were homozygous for disrupted FAD2 alleles. In the allohexaploid, Camelina, guide RNAs were designed that simultaneously targeted all three homoeologous FAD2 genes. This strategy that significantly enhanced oil composition in T₃ and T₄ generation Camelina seeds was associated with a combination of germ‐line mutations and somatic cell mutations in FAD2 genes in each of the three Camelina subgenomes.     

A toolkit for Nannochloropsis oceanica CCMP1779 enables gene stacking and genetic engineering of the eicosapentaenoic acid pathway for enhanced long‐chain polyunsaturated fatty acid production

Nannochloropsis oceanica is an oleaginous microalga rich in ω3 long‐chain polyunsaturated fatty acids (LC‐PUFAs) content, in the form of eicosapentaenoic acid (EPA). We identified the enzymes involved in LC‐PUFA biosynthesis in N. oceanica CCMP1779 and generated multigene expression vectors aiming at increasing LC‐PUFA content in vivo. We isolated the cDNAs encoding four fatty acid desaturases (FAD) and determined their function by heterologous expression in S. cerevisiae. To increase the expression of multiple fatty acid desaturases in N. oceanica CCMP1779, we developed a genetic engineering toolkit that includes an endogenous bidirectional promoter and optimized peptide bond skipping 2A peptides. The toolkit also includes multiple epitopes for tagged fusion protein production and two antibiotic resistance genes. We applied this toolkit, towards building a gene stacking system for N. oceanica that consists of two vector series, pNOC‐OX and pNOC‐stacked. These tools for genetic engineering were employed to test the effects of the overproduction of one, two or three desaturase‐encoding cDNAs in N. oceanica CCMP1779 and prove the feasibility of gene stacking in this genetically tractable oleaginous microalga. All FAD overexpressing lines had considerable increases in the proportion of LC‐PUFAs, with the overexpression of Δ12 and Δ5 FAD encoding sequences leading to an increase in the final ω3 product, EPA.