The Effect of Phytohormones on Chlorophyll(ide), Protochlorophyll(ide) and Carotenoid Formation in Greening Dark Grown Wheat Leaves

The influence of phytohormones on chlorophyll and carotenoid formation during the greening of irradiated dark grown wheat leaves (Triticum aestivum L. cv. Starke II Weibull) was studied. Leaves were floated on solutions of abscisic acid, gibberellic acid and kinetin for 24 h. The chlorophyll and carotenoid contents were determined during a subsequent period of 48 h of continuous irradiation. Leaves treated with abscisic acid showed a longer lag phase and a lower rate of accumulation of chlorophyll as compared to the control than did leaves treated with gibberellic acid and kinetin. The carotenoid content was low both in leaves treated with abscisic acid and in those treated with gibberellic acid. Treatment with abscisic acid lowered the protochlorophyllide regeneration after a saturating light flash while gibberellic acid as well as kinetin had no effect. The influence of ABA was partly dependent on an increase of the wounded part of the cut leaf segments. The accumulation of protochlorophyllide in leaves treated with δ-aminolevulinic acid was not affected by the different hormonal treatments. These results suggest that the main effect of abscisic acid is probably outside the chloroplast, i.e. on the formation or transport of δ-aminolevulinic acid.     

Effect of osmotic stress and sodium nitroprusside pretreatment on proline metabolism of wheat seedlings

Effect of osmotic stress and sodium nitroprusside (SNP, NO donor) pretreatment on growth and proline metabolism of wheat seedlings was investigated. Polyethylene glycol 6000 treatment for 2, 4 and 6 d could be termed as mild, moderate and severe stress, respectively, according to decrease in the relative water content. Severe osmotic stress significantly decreased the growth and photochemical efficiency, and increased proline content due to activation of its synthesis. 0.2 mM SNP pretreatment enhanced growth of wheat seedlings, increased variable to maximum fluorescence ratio (F_v/F_m) and fluorescence yield, while decreased proline content. However, 2 mM SNP retarded the seedlings growth and chlorophyll a fluorescence, and increased proline accumulation. Our results showed that NO might be involved in the regulation of osmotic stress in a concentration-dependent manner.     

The effect of plant-hormone pretreatments on ethylene production and synthesis of 1-aminocyclopropane-1-carboxylic acid in water-stressed wheat leaves

Excised wheat (Triticum aestivum L.) leaves, when subjected to drought stress, increased ethylene production as a result of an increased synthesis of 1-aminocyclopropane-1-carboxylic acid (ACC) and an increased activity of the ethyleneforming enzyme (EFE), which catalyzes the conversion of ACC to ethylene. The rise in EFE activity was maximal within 2 h after the stress period, while rehydration to relieve water stress reduced EFE activity within 3 h to levels similar to those in nonstressed tissue. Pretreatment of the leaves with benzyladenine or indole-3-acetic acid prior to water stress caused further increase in ethylene production and in endogenous ACC level. Conversely, pretreatment of wheat leaves with abscisic acid reduced ethylene production to levels produced by nonstressed leaves; this reduction in ethylene production was accompanied by a decrease in ACC content. However, none of these hormone pretreatments significantly affected the EFE level in stressed or nonstressed leaves. These data indicate that the plant hormones participate in regulation of water-stress ethylene production primarily by modulating the level of ACC.Abbreviations ABA abscisic acid - ACC 1-aminocyclopropane-1-carboxylic acid - BA N⁶-benzyladenine - EFE ethylene-forming enzyme - IAA indole-3-acetic acid     

外源脯氨酸对高温胁迫下黄瓜幼苗叶片AsA-GSH循环和光合荧光特性的影响

以抗热性较弱的黄瓜品种‘新泰密刺'为试材,在人工气候箱内采用营养液栽培法,研究了外源脯氨酸(Pro)预处理对高温胁迫下黄瓜幼苗叶片抗坏血酸-谷胱甘肽循环和光合荧光特性的影响.结果显示:(1)与清水处理相比,高温胁迫4 h和8 h时,Pro预处理黄瓜幼苗叶片单脱氢抗坏血酸还原酶(MDAR)活性、GSH(还原型谷胱甘肽)/GSSG(氧化型谷胱甘肽)比值及GSH含量显著升高;(2)在高温胁迫8 h时,Pro预处理幼苗的净光合速率(P_n)、气孔导度(G_s)及PSⅡ的最大光化学效率(F_v/F_m)、光化学淬灭系数(q_P)均显著升高,而蒸腾速率(T_r)和非光化学淬灭系数(NPQ)降低.研究表明,外源Pro预处理可显著提高高温胁迫下黄瓜幼苗叶片抗坏血酸-谷胱甘肽循环清除H_2O_2能力和叶片光合能力,有效缓解高温胁迫对黄瓜叶片抗氧化系统和光合系统的伤害,从而增强植株的耐热性.     

Hydrogen peroxide pretreatment induces osmotic stress tolerance by influencing osmolyte and abscisic acid levels in maize leaves

Hydrogen peroxide (H₂O₂) functions as a signal molecule in plants under abiotic and biotic stresses. Leaves of detached maize (Zea mays L.) seedlings were used to study the function of H₂O₂ pretreatment in osmotic stress resistance. Low H₂O₂ concentration (10 mM) which did not cause a visual symptom of water deficit (leaf rolling) was applied to the seedlings. Exogenous H₂O₂ alone increased leaf water potential, endogenous H₂O₂ content, abscisic acid (ABA) concentration, and metabolite levels including soluble sugars, proline, and polyamines while it decreased lipid peroxidation and stomatal conductance. Osmotic stress induced by polyethylene glycol (PEG 6000) decreased leaf water potential and stomatal conductance but enhanced lipid peroxidation, endogenous H₂O₂ content, the metabolite levels, and ABA content. H₂O₂ pretreatment also induced the metabolite accumulation and improved water status, stomatal conductance, lipid peroxidation, ABA, and H₂O₂ levels under osmotic stress. These results indicated that H₂O₂ pretreatment may alleviate water loss and induce osmotic stress resistance by increasing the levels of soluble sugars, proline, and polyamines thus ABA and H₂O₂ production slightly decrease in maize seedlings under osmotic stress.     

The effect of water deficit and excess copper on proline metabolism in Nicotiana benthamiana

Fluctuation in proline content is a widespread phenomenon among plants in response to heavy metal stress. To distinguish between the participation of water deficit and copper on changes in proline metabolism, potted plants and floating leaf discs of tobacco were subjected to CuSO₄ treatments. The application of copper increased the proline content in the leaves concomitantly with decreased leaf relative water content and increased abscisic acid (ABA) content in the potted plant. Excess copper increased the expression of two proline synthesis genes, pyrroline-5-carboxylate synthetase (P5CS) and ornithine aminotransferase (OAT) and suppressed proline catabolism gene, proline dehydrogenase (PDH). However, in the experiment with tobacco leaf discs floating on CuSO₄ solutions, the excess copper decreased proline content and suppressed the expression of the P5CS, OAT and PDH genes. Therefore, proline accumulation in the potted tobacco plants treated with excess Cu treatment might not be the consequence of the increased copper content in tobacco leaves but rather by the accompanied decrease in water content and/or increased ABA content.     

The effect of simulated microgravity on formation of the pigment apparatus in etiolated barley seedlings

The effect of horizontal clinorotation on the dynamics of the accumulation of the main photosynthetic pigments in the greening of 6-day-old etiolated barley seedlings has been studied. The content of protochlorophillide, the direct precursor of chlorophyll a, in clinorotated seedlings in the dark was 9–20% lower than in the control group. After exposure of barley seedlings to light for 12 h under clinorotation, chlorophyll accumulation lagged behind the control by 45% and reached the control value after 48–72 h. The total content of carotenoids increased many fold during greening; at the first stage the carotenoid level in clinorotated seedlings was less than in the control. The synthesis rates of δ-aminolevulinic acid and δ-aminolevulinate dehydratase activity in clinorotated seedlings were slower than in the control after 24 h of greening and after 72 h of greening reaching the control values. The activity of Mg-protoporphyrin IX chelatase catalyzing the incorporation of Mg ions in the structure of chlorophyll a, did not change when exposed to clinorotation. The results we obtained show inhibition of the initial stages of chlorophyll biosynthesis in the conditions of simulated microgravity. The light, to a certain extent, decreases the negative effect of microgravity on the formation of the photosynthetic apparatus in plants.     

The Relationship between Chlorophyllide Accumulation, the Amount of Protochlorophyllide636 and Protochlorophyllide650 in Dark Grown Wheat Leaves Treated with δ-Aminolevulinic Acid

The influence of different amounts of protochlorophyl-lide₆₃₆ and protochlorophyllide₃₅₀ on light Induced chloro-phyllide formation was. studied in wheat leaves treated with δ-aminolevulinic acid. The phototransformation of proto-chlorophyllide was performed with weak red light. This transformation is unaffected by the δ-aminolevulinic acid treatment, whilst the accumulation of chlorophyllide, both the rate and the amount, is greatly stimulated by moderate amounts of protochlorophyllide₆₃₆. The presence of large amounts of protochlorophyllide₆₃₆ decreases the rate of chlorophyllide formation, but increases the final amount of chloro-phyllide formed. A decreased level of protochlorophyllide₆₅₀, obtained by treatment with N_aN₃, results in a decreased transformation rate. Inhibitors; of protein synthesis do not seem to influence the transformation of protochlorophyllide₆₃₆ to chlorophyllide, suggesting that no new synthesis of protein is required. The experimental results indicate that the final steps in chlorophyll biosynthesis are protochlorpnyllide₆₃₆→ protochlorophyllide₆₅₀→ chlorophyllide → chlorophyll.     

Effects of Kinetin on Transpiration Rate and Abscisic Acid Content of Water Stressed Young Wheat Plants

Effects of kinetin on transpiration rate and abscisic acid content were determined. Leaves from 9-day-old wheat plants (Triticum aestivum L. cv. Weibull's Starke II) were used. —Transpiration rate decreased in excised leaves put in water, but it was maintained at a higher rate when kinetin was supplied. When excised leaves were water stressed by air-drying for 1 h, addition of kinetin resulted in a considerable stimulation of transpiration rate. The effect reached its maximum after 15 h and this level remained relatively unchanged for at least 10 h. Intact seedlings which were stressed before leaf excision, showed only a slight stimulation of kinetin on transpiration rate. — Abscisic acid content slowly increased up to three-fold in 2 days in excised leaves put in water. In excised and water-stressed leaves the abscisic acid content was reduced during the first 24 h and then increased. As the leaves were fully turgid, the increase could not have been caused by water stress. However, both in stressed and unstressed leaves kinetin addition reduced the increase in abscisic acid content. — It is suggested that the stimulation by kinetin on transpiration rate in excised and water stressed leaves was mainly due to the combined effect of (1) a reduction in the activity of endogenous cytokinins, (2) kinetin acting as a ‘substitute’ for the inactivated cytokinins but exerting a stronger effect on transpiration than the endogenous cytokinins, and (3) the ‘extra’ reduction in abscisic acid content caused by the kinetin treatment. Furthermore, the results indicate that changes in cytokinins might be partly responsible for the aftereffect on transpiration.     

Influence of water stress on endogenous hormone contents and cell damage of maize seedlings

Phytohormones play critical roles In regulating plant responses to stress. We Investigated the effects of water stress Induced by adding 12% (w/v) polyethylene glycol to the root medium on the levels of abscisic acid (ABA), indole-3-acid (IAA), zeatin (ZT), and gibberellin3 (GA3) in maize leaves. The results suggested that water stress had significant effects on the four hormone levels. There was a transient increase in the IAA content during the initial stage of adaptation to water stress in maize leaves, but it dropped sharply thereafter in response to water stress. ABA content increased dramatically in maize leaves after 24 h of exposure to water stress, and then the high levels of ABA were maintained to the end, The contents Of ZT and GA3 rapidly declined in maize leaves subjected to water stress. The effects of water stress on chlorophyll content, electrolyte leakage and malondialdehyde levels in maize leaves were also studied. The variation of cell damage was negatively correlated with ZT and GA3 levels in maize leaves under water stress. Thus, we explored the roles of ZT and GA3 on the growth of maize seedlings under water stress by exogenous application. It is possible that both ZT and GA3 were effective in protecting maize seedlings from water stress, which would be of great importance for the improvement of drought tolerance in maize by genetic manipulation.     

干旱胁迫对不同土壤基质下核桃楸幼苗的生理特性的影响

为探究不同土壤基质条件下核桃楸幼苗对干旱胁迫生理特性的响应,以两年生核桃楸幼苗为研究对象,在腐殖土、壤土和砂壤土三种基质条件下进行控水试验,水分梯度分别设置为CK(正常供水,80%田间持水量)、T1(轻度胁迫,60%田间持水量)、T2(中度胁迫,40%田间持水量)和T3(重度胁迫,20%田间持水量),测定幼苗叶片叶绿素、丙二醛、脯氨酸、抗氧化酶和光合指标。结果表明,随着胁迫程度的加深,叶绿素含量和Chla/b均呈逐渐降低的趋势(P<0.05),在胁迫60d时,T3处理下的叶绿素a(Chla)和叶绿素b(Chlb)分别比CK低65.41%和51.57%,3种基质中叶绿含量大小顺序为:腐殖土>壤土>砂壤土。随着胁迫程度的加深,叶片脯氨酸(Pro)、丙二醛(MDA)则呈逐渐升高的趋势(P<0.05),胁迫60d时,砂壤土中T3处理的Pro含量最高为83.37mg·g-1,比CK高86.32%。腐殖土中幼苗叶片超氧化物歧化酶(SOD)、过氧化物酶(POD)和过氧化氢酶(CAT)活性均随胁迫程度加深而升高,壤土中叶片SOD活性则呈先升高后降低的变化规律。3种土壤基质中核桃楸叶片净光合速率(Pn)、蒸腾速率(Tr)和气孔导度(Gs)均随着胁迫程度的加深而降低,但不同基质中幼苗叶片的气孔限制和非气孔限制发生的时间不同,T3处理的叶片胞间CO2浓度(Ci)在腐殖土中是12:00时最低,而在壤土和砂壤土中则是10:00时最低。土壤全氮、全磷、渗透速率和蒸腾速率与叶绿素含量存在显著的正相关关系(P<0.05),土壤容重与叶绿素含量存在显著的负相关关系(P<0.05)。因此,土壤基质和干旱胁迫对核桃楸幼苗的生理生化和光合特性产生了显著的影响,重度胁迫对幼苗的叶绿素、渗透物质和酶体系产生了严重的损伤,同时降低了核桃楸幼苗的光合生产力。总体上,腐殖土中的幼苗有着较好的抗旱表现。     

Photosynthetic responses and proline content of mature and young leaves of sunflower plants under water deficit

In mature and young leaves of sunflower (Helianthus annuus L. cv. Catissol-01) plants grown in the greenhouse, photosynthetic rate, stomatal conductance, and transpiration rate declined during water stress independently of leaf age and recovered after 24-h rehydration. The intercellular CO₂ concentration, chlorophyll (Chl) content, and photochemical activity were not affected by water stress. However, non-photochemical quenching increased in mature stressed leaves. Rehydration recovered the levels of non-photochemical quenching and increased the F_v/F_m in young leaves. Drought did not alter the total Chl content. However, the accumulation of proline under drought was dependent on leaf age: higher content of proline was found in young leaves. After 24 h of rehydration the content of proline returned to the same contents as in control plants.     

The Effect of Polyethylene Glycol on Proline Accumulation in Rice Leaves

The regulation of proline accumulation in polyethylene glycol (PEG, –1.5 MPa) treated rice leaves was investigated. PEG treatment resulted in a decrease in relative water content, indicating that PEG treatment caused water stress in rice leaves. Proline accumulation caused by PEG was related to protein hydrolysis, an increase in ornithine--amino- transferase activity, an increase in the content of ammonia, and an increase in the contents of the precursors of proline biosynthesis, glutamic acid, ornithine, and arginine. Results also show that abscisic acid accumulation is not required for proline accumulation in PEG-treated rice leaves.     

NaCl pretreatment alleviates salt stress by enhancement of antioxidant defense system and osmolyte accumulation in mungbean (Vigna radiata L. Wilczek)

Enhancement of salt (NaCl) tolerance by pretreatment with sublethal dose (50 mM) of NaCl was investigated in V. radiata seedlings. NaCl stress caused drastic effects on roots compared to shoots. Accompanying reductions in length, number of root hairs and branches, roots became stout, brittle and brown in color. Salt stress caused gradual reduction in chlorophyll, carotenoid pigment contents and chlorophyll fluorescence intensity also. Superoxide dismutase and catechol peroxidase activities increased under stress in both roots and leaves. But catalase activity showed an increase in roots and decrease in leaves. In these seedlings, the oxidative stress has been observed under salinity stress and the level of proline, H2O2 and malondialdehyde content were increased. But pretreatment with sublethal dose of NaCl was able to overcome the adverse effects of stress imposed by NaCl to variable extents by increasing growth and photosynthetic pigments of the seedlings, modifying the activities of antioxidant enzymes, reducing malondialdehyde and H2O2 content and increasing accumulation of osmolytes like proline. Thus, mungbean plants can acclimate to lethal level of salinity by pretreatment with sublethal level of NaCl, improving their health and production under saline condition.     

Physiological Evaluation of Drought Stress Tolerance and Recovery in Cauliflower (Brassica oleracea L.) Seedlings Treated with Methyl Jasmonate and Coronatine

Coronatine (COR) is a chlorosis-inducing phytotoxin that mimics some biological activities of methyl jasmonate (MeJA). Although MeJA has been reported to alleviate drought stress, it is unclear if COR has the same ability. Our objective was to determine the influence of exogenously applied MeJA and COR on the growth and metabolism of cauliflower seedlings under drought stress and recovery. Both MeJA and COR enhanced the growth and accumulation of dry matter in cauliflower seedlings during drought-stressed and rewatering conditions. Treatment with MeJA or COR enhanced tolerance of drought stress through increased accumulation of chlorophyll and net photosynthetic rate. Enzymatic (superoxide dismutase, peroxidase, catalase, ascorbate peroxidase, and glutathione reductase) and nonenzymatic antioxidant (proline and soluble sugar) systems were activated, and lipid peroxidant (malondialdehyde and hydrogen peroxide) was suppressed by MeJA and COR under drought stress. MeJA and COR also increased leaf relative water content and endogenous abscisic acid level under drought-stressed conditions. After rewatering, the contents of leaf water, chlorophyll, abscisic acid, and photosynthetic characteristics as well as enzymatic and nonenzymatic antioxidant systems showed nearly complete recovery. Both MeJA and COR can alleviate the adverse effects of drought stress and enhance the ability for water stress resistance through promotion of defense-related metabolism in cauliflower seedlings.     

栓皮栎幼苗对土壤干旱胁迫的生理响应

以栓皮栎一年生盆栽苗为实验材料,采用称重控水的方法,设置不同土壤水分胁迫梯度,系统分析其幼苗在不同干旱胁迫条件下的生理生化响应特征,以探索栓皮栎耐旱特性.结果显示:(1)栓皮栎幼苗叶片中3种保护酶(SOD、POD、CAT)活性在对照(CK,土壤相对含水量19.5％～21.5％)条件下保持稳定,而中度干旱(T2,9.5％～11.5％)和重度干旱(T3,5.5％～7.5％)条件下,随着胁迫时间的延长呈先增高后降低的趋势,且变化的幅度在不同胁迫强度下存在差异.(2)在整个干旱胁迫过程中,各胁迫处理叶片丙二醛(MDA)含量均呈上升趋势,不同胁迫强度的变化幅度不同;叶片中的可溶性蛋白含量和根系活力随着干旱胁迫程度的增强呈先增高后降低的趋势.(3)栓皮栎幼苗叶片的脯氨酸含量随着干旱胁迫时间的延长表现出先增加后降低的趋势;叶片叶绿素a、叶绿素b、总叶绿素含量以及叶绿素a/b值均呈逐渐降低的趋势.研究表明,栓皮栎幼苗在短期和轻度干旱胁迫下通过提高自身的保护酶活性、增加可溶性蛋白和脯氨酸含量、提高根系活力等来抵御干旱环境的伤害,从而表现出较强的耐旱特性;而在重度干旱胁迫条件下,栓皮栎幼苗自我调节能力丧失,体内代谢紊乱,导致保护酶活性、可溶性蛋白、脯氨酸含量和根系活力等下降,从而受到干旱伤害.     

Effect of nitrogen deficiency on antioxidant status and Cd toxicity in rice seedlings

Effect of nitrogen (N) deficiency on antioxidant status and Cd toxicity in rice seedlings was investigated. N deficiency resulted in a reduction of shoot growth but not root growth. The contents of N-containing compounds such as nitrate, chlorophyll, and protein decreased in leaves of rice seedlings grown under N deficiency. Accumulation of abscisic acid and H₂O₂ in leaves was induced by N deficiency. The content of ascorbate and the activities of ascorbate peroxidase, glutathione reductase, and catalase in N-deficient leaves were lower than their respective control leaves. However, glutathione content was not affected and superoxide dismutase activity was increased by N deficiency. Cd toxicity in N-deficient seedlings was more pronounced than that in N-sufficient ones. Pretreatment with ascorbate or L-galactono-1,4-lactone, a biosynthetic precursor of ascorbate resulted in a reduction of Cd toxicity enhanced by N deficiency. N deficiency also resulted in an enhancement of Cd uptake in rice seedlings. The possible mechanism of Cd toxicity enhanced by N deficiency is discussed.     

外源硫化氢和水杨酸对盐胁迫下加工番茄幼苗生长与生理特性的影响

以加工番茄KT-7为材料,在水培条件下,研究外源水杨酸(SA,0.15 mmol/L)、硫化氢(H2S)供体硫氢化钠(NaHS,50 mmol/L)对150 mmol/L NaCl胁迫下加工番茄幼苗的渗透调节、活性氧代谢和快速叶绿素荧光的影响,以探讨H2S和SA这2种信号分子协同作用、以及实际生产中缓解加工番茄幼苗盐胁...     

Implication of ABA and proline on cell membrane injury of water deficit stressed barley seedlings

The aim of this work was to examine the ability of ABA and proline to counteract the deleterious effect of water deficit stress on cell membrane injuries. Six-day-old seedlings of two barley genotypes (cv. Aramir, line R567) were treated with ABA (2·10⁻⁴ M) or proline (0.1 M) for 24 h, and then subjected to osmotic stress for 24h, by immersing their roots in polyethylene glycol (PEG 6000) solution of osmotic potential of −1.0 MPa and −1.5 MPa or by submerging the leaf pieces in PEG solution of osmotic potential of −1.6 MPa. Pretreatment of plants with ABA and proline caused an increase of free proline level in the leaves. Plants treated with ABA exhibited a lower membrane injury index under water stress conditions than those untreated even when no effect of this hormone on RWC in the leaves of stressed plants was observed. Pretreatment of plants with proline prevented to some extent membrane damage in leaves of the stressed seedlings, but only in the case when stress was imposed to roots. Improvement in water status of leaves was also observed in seedlings pretreatment with proline. The protective effect of both ABA and proline was more pronounced in line R567 that exhibited higher membrane injury under water deficit stress conditions.     

黄腐酸和甜菜碱预处理对干旱胁迫下平邑甜茶生理特性及光合的影响

该研究以平邑甜茶[Malus hupehensis(Pamp.)Rehd.]2年生实生苗为材料,通过盆栽试验于干旱处理前3d分别连续喷施黄腐酸(FA)、甜菜碱(GB)和复配(FA+GB),并以清水为对照(CK)进行预处理,比较分析不同预处理对干旱胁迫下平邑甜茶的生理及光合特性变化,探讨FA和GB对平邑甜茶的抗旱生理机制。结果显示:(1)与对照相比,FA、GB和FA+GB预处理均能够显著提高平邑甜茶叶片相对含水量,且FA的保水性效果最佳。(2)3种预处理均可显著促进干旱胁迫下叶片可溶性蛋白、可溶性糖和脯氨酸含量增加,且FA+GB预处理后在干旱胁迫下叶片可溶性糖和脯氨酸累积量显著高于单施FA或GB。(3)3种预处理均可显著提高干旱胁迫下平邑甜茶幼苗的SOD、POD、CAT活性,并显著降低MDA的积累速度及其累积量,且以FA+GB预处理的MDA含量最低、抗氧化酶活性最高。(4)GB和FA+GB预处理下平邑甜茶的净光合速率、瞬时水分利用率显著高于CK和FA,且FA+GB处理下改善光合特性的效果最佳,GB次之。研究表明,单独喷施黄腐酸和甜菜碱及两者配施预处理均能够增加干旱胁迫下平邑甜茶的渗透调节物质和相对含水量,提高叶片的保水性,调节抗氧化物酶活性,降低丙二醛含量,增加细胞膜稳定性,改善光合性能,进而提高平邑甜茶的抗旱能力,且以复配喷施(FA+GB)预处理的效果最好。