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1.
Leonardo Schena Franco Nigro Isabella Pentimone Angela Ligorio Antonio Ippolito 《Postharvest Biology and Technology》2003,30(3):209-220
Fifty-one endophytic isolates of Aureobasidium pullulans were obtained from the flesh of sweet cherries and extensively screened to evaluate their biocontrol activity against postharvest rots of sweet cherries and table grapes. Preliminary analysis of all isolates by randomly amplified polymorphic DNA (RAPD) with three different primers showed the presence of a high genetic variability and enabled isolates not showing any genetic difference to be discarded. Thirty-five isolates with different RAPD electrophoresis patterns had a wide range of biocontrol activity against Botrytis cinerea and Monilinia laxa on single-wounded berries of sweet cherries and table grapes with a reduction of decay from 10 to 100%. Two isolates (533 and 547) significantly reduced B. cinerea on table grape berries also when applied 6, 12, and 24 h after the pathogen inoculation. In a 2-year period of investigation (1998–1999), a reduction of total rots ranging from 32 to 80% (sweet cherries) and from 59 to 64% (table grape) was achieved with isolates 533 and 547 applied after harvest. Preharvest applications of isolate 547 significantly reduced postharvest rots of sweet cherries and table grapes by 47 and 38%, respectively. On the whole, isolates 533 and 547 were more effective than A. pullulans L47, a biocontrol agent of postharvest diseases with a known activity. Population studies demonstrated that isolate 547 was able to survive under field conditions, to increase its population during cold storage, and to penetrate the flesh of sweet cherries when applied during flowering. 相似文献
2.
Giuseppe Lima Filippo De Curtis Daniela Piedimonte Anna Maria Spina Vincenzo De Cicco 《Postharvest Biology and Technology》2006,40(3):301-307
Three selected biocontrol yeasts, known for their elevated antagonistic activity against several fungal pathogens, were tested in vitro for compatibility with common fungicides. The antagonists were resistant to several fungicides, but they were inhibited by triazoles and dithiocarbamates. The yeast isolate LS28 (Cryptococcus laurentii), which tolerated in vitro high rates of benzimidazoles, was tested, alone or in combination with a low dose (10% of the full label rate) of thiabendazole, against grey mould on stored apples. Wounded fruit were inoculated with thiabendazole resistant and sensitive isolates of Botrytis cinerea, applied separately or as a mixture. A more effective and prolonged control of fungal decay was always exerted by the biocontrol yeast applied together with thiabendazole at a low dose. This treatment provided synergistic effects and was markedly better than treatments applied separately, whereas the fungicide applied alone at the highest label dose was ineffective in the presence of the isolate of B. cinerea resistant to thiabendazole. The biocontrol yeast applied alone provided significant protection of apples stored for up to 12 days at 20 °C (from 39 to 81% less decay than untreated control), whereas the fungicide alone at low or high dose was always ineffective in the presence of the B. cinerea isolate resistant to benzimidazoles. The integrated treatment was highly effective and durable showing high reduction of decay (from 66 to 92%) even after 18 days of storage. This study suggests that the integration of a biocontrol agent with a low rate of fungicide may be a useful alternative strategy to manage both sensitive and resistant isolates of fungal pathogens efficiently and to reduce risks involved with extensive use of synthetic chemicals. 相似文献
3.
Postharvest decay of pear fruit often originates at small wounds that occur during harvest and handling. Experiments were conducted to characterize the effect of timing of application of postharvest decay control materials, and to evaluate sequential postharvest applications of fungicides or fungicides and biocontrol agents. Fungicides and biocontrol agents were increasingly less effective when the period between harvest and application was prolonged. Thiabendazole (TBZ) applied to fruit without artificial wounding or inoculation effectively reduced decay when applied within 3 weeks or 6 weeks in 2 years of study. TBZ, fludioxonil, and pyrimethanil were effective in controlling decay at artificial wounds inoculated with Penicillium expansum up to 14 d after inoculation. Application of TBZ at harvest followed 3 weeks later by application of fludioxonil was superior to application of TBZ at harvest alone. Three yeast and one bacterial biocontrol agents reduced decay at pear wounds inoculated with P. expansum up to 14 d after inoculation with P. expansum, but were ineffective when applied at 28 d after inoculation. Of possible sequential arrangements of fungicide and biocontrol treatments, application of the most effective material promptly after harvest generally resulted in the highest level of decay control. 相似文献
4.
Yan Zhao Kang Tu Xingfeng Shao Wei Jing Zipeng Su 《Postharvest Biology and Technology》2008,49(1):113-120
The yeast Pichia guilliermondii was examined for its ability to control Rhizopus nigricans on tomato fruit during storage, and in order to highlight the reason for biocontrol, a possible mode of action is discussed. Results showed that autoclaved yeast culture and culture filtrate had no effect on controlling the postharvest disease caused by R. nigricans, although inoculation of P. guilliermondii prior to R. nigricans resulted in enhanced biocontrol efficacy. Moreover, rapid colonization of the yeast on wound sites was observed during the initial 3 days at 20 °C, and then the population stabilized for the remaining 4 days. This phenomenon indicated that at room temperature, P. guilliermondii could acclimatize itself to the environment of tomato fruit wounds and occupy the living space quickly. The results indicate that P. guilliermondii did not produce an antifungal substance, however, competition for nutrients and space on wounds appeared to play a role in the activity of the biocontrol and could be one of the mechanisms. In addition, the fruit inoculated with P. guilliermondii demonstrated changes in peroxidase (POD), polyphenoloxidase (PPO), superoxide dismutase (SOD), catalase (CAT), phenylalanine ammonia-lyase (PAL), chitinase (CHI) and β-1,3-glucanase activities, all of which were correlated with the onset of induced resistance. This result suggests that tomato fruit is capable of responding to the yeast P. guilliermondii, which could activate defensive enzymes and thereby induce host disease resistance. 相似文献
5.
Teresa P. Caams Immaculada Vias Josep Usall Rosario Torres Marina Anguera Neus Teixid 《Postharvest Biology and Technology》2008,49(1):96-106
Infection of citrus fruit by postharvest pathogens often occurs in the field prior to harvest; therefore, it could be advantageous to apply biocontrol agents before harvest, which would reduce initial infection and then remain active and control pathogens in storage and under commercial conditions. The objective of the present work was to evaluate the effectiveness of different formulations of Pantoea agglomerans applied preharvest for controlling postharvest diseases on citrus. Results confirmed the protective effect of the additive Fungicover (FC) on populations of P. agglomerans exposed to non-conducive field conditions. In general, when osmotic-adapted and lyophilised P. agglomerans cells were used in bacterial treatments, these treatments showed greater survival rates than treatments with non-osmotic-adapted or fresh cells under field conditions. However, this superiority was only found when Fungicover was also added to suspensions of bacterial treatments. Therefore, bacterial treatments with Fungicover had population levels of P. agglomerans cells 1.2 and 2.8 log CFU cm−2 higher than bacterial treatments without Fungicover during field experiments. These results allowed us to conclude that it is possible to improve environmental stress tolerance and ecological competence of P. agglomerans cells by integrating certain formulation strategies. Consequently, the improved formulation of P. agglomerans provided an effective control for orange fruit against natural postharvest pathogen infections and artificial infections of Penicillium digitatum with values of decay reduction higher than 50%. These latter results also demonstrated that it is possible to control postharvest pathogens using bacterial preharvest treatments. 相似文献
6.
为研究扑草净对10种小麦田恶性杂草的防除活性,通过温室整株生物测定法比较研究扑草净对菵草(Beckmannia syzigachne)、看麦娘(Alopecurus aequalis)、日本看麦娘(Alopecurus japonicus)、大穗看麦娘(Alopecurus myosuroides)、多花黑麦草(Lolium multiflorum)、耿氏假硬草(Pseudosclerochloa kengiana)、早熟禾(Poa annua)、播娘蒿(Descurainia sophia)、猪殃殃(Galium spurium)、荠菜(Capsella bursa-pastoris)等10种小麦田恶性杂草土壤处理和早期茎叶处理（禾本科杂草2~3叶期,阔叶杂草2对或轮叶期）的防除活性。结果表明,扑草净推荐剂量（有效成分750 g/hm2）早期茎叶处理后30天对猪殃殃、播娘蒿、荠菜的地上部分鲜重抑制率分别达89.75%、94.04%、75.00%,推荐剂量1/2量对播娘蒿2对叶龄幼苗地上部分鲜重抑制率达到89.64%。扑草净推荐剂量早期茎叶处理后30天完全杀灭看麦娘、日本看麦娘、耿氏假硬草,对大穗看麦娘、早熟禾、菵草、多花黑麦草地上部分鲜重抑制率分别为95.80%、83.58%、82.77%、60.11%。扑草净推荐剂量土壤处理后30天对看麦娘、耿氏假硬草、早熟禾、菵草的地上部分鲜重抑制率达到85%以上。扑草净早期茎叶处理对看麦娘、日本看麦娘、大穗看麦娘、耿氏假硬草、早熟禾、播娘蒿、猪殃殃有效,对菵草和荠菜也有一定的防除活性,对多花黑麦草防效差。 相似文献
7.
番茄产后灰霉病的病原鉴定及生物防治 总被引：1，自引：1，他引：0
为探索生物防治番茄产后灰霉病,采用柯赫氏法则从罹病的番茄果实上分离得到菌株FQ-2,基于形态学和rDNA-ITS序列,对该菌株进行病原鉴定,采用小麦内生菌解淀粉芽孢杆菌 B9601-Y2(Y2)和球花石斛内生菌贝莱斯芽孢杆菌DJB5(DJB5)对番茄灰霉病进行生物防治。FQ-2被鉴定为灰葡萄孢菌(Botrytis cinerea),且Y2和DJB5对番茄产后果实灰霉病均有较好的预防和治疗效果,Y2 1×107 cfu/mL和DJB5 1×108 cfu/mL悬浮剂在处理当天接种FQ-2,10天后的防效分别达88.10%和73.47%。在接种灰霉病菌当天再接种1×108cfu/mL的Y2和1×107 cfu/mL的DJB5,10天后灰霉病的防治效果分别达68.56%和78.74%。研究结果对番茄产后灰霉病的生物防治具有参考作用。 相似文献
8.
拟康氏木霉和黑根霉混合发酵对苹果链格孢的防治 总被引：2，自引：0，他引：2
研究黑根霉和拟康氏木霉混合发酵液对苹果链格孢菌的生物防治效果,探索开发为新型绿色环保生物防治剂。对2种生防微生物进行形态学鉴定和ITS鉴定;生长速率法测定混合发酵液的抑菌活性;在烟台莱阳进行田间实验对发病较为严重的果园进行为期一年的多次灌根与喷施处理,测定其防治效果并测定其对苹果叶片SOD、POD等防御相关酶的活性。研究结果表明,黑根霉和拟康氏木霉混合发酵可显著提高对病原微生物链格孢菌的抑菌能力。平板抑菌实验结果表明,混合发酵液对链格孢菌的抑菌效果达到了66.73%,显著高于黑根霉和拟康氏木霉单独发酵液的51.23%、17.28%;田间实验发现,混合发酵液处理后苹果斑点落叶病发病率与对照组相比显著下降,相对防效达到42.46%;对其叶片防御酶测定结果表明,混合发酵液处理后,苹果叶片的防御酶SOD、POD、PAL活性均有50%以上的提高。黑根霉和拟康氏木霉混合发酵液对苹果链格孢菌具有显著的抑制作用,能有效防治苹果斑点落叶病,提高苹果抗病能力,为新型生防制剂的开发提供了参考。 相似文献
9.
为明确内生解淀粉芽孢杆菌(Bacillus amyloliquefaciens)菌株P-r21的抑菌广谱性及对番茄灰霉病的防治效果,本研究优化了菌株P-r21发酵条件,利用平板对峙法测定了该菌株对6种病原菌的抑菌活性及对番茄灰霉病菌(Botrytis cinerea)孢子萌发的抑制作用,并通过离体接种试验研究了对番茄灰霉病的防效。结果表明：菌株P-r21最佳发酵条件为在PYJ中培养初始pH 6.0,培养时间为12 h。菌株P-r21对供试的6种植物病原菌(Botrytis cinerea、Fusarium proliferatum、Rhizoctonia cerealis、Gaeumannomyces graminis、Fusarium graminearum、Colletotrichum karstii)的生长均表现出抑制作用;对C. karstii和B.cinerea抑制作用最强,抑菌率达到88.0%和84.5%。发酵滤液对B. cinerea孢子萌发抑制率达80.6%,对叶片和果实灰霉病的防效达到100%和80.0%;发酵液10倍稀释后防效也可达到67.1%和60.7%。本研究明... 相似文献
10.
为评估产紫青霉对烟草根结线虫病的生防潜质,采用产紫青霉K1对南方根结线虫二龄幼虫及卵的活性进行了室内生测,同时利用了产紫青霉K1生物菌剂对烟草根结线虫病进行了2年2地的田间防效测定。结果表明：（1）产紫青霉K1菌株对南方根结线虫具有较强的毒杀活性,48 h对根结线虫2龄幼虫的校正致死率高达94.53%,对卵孵化的相对抑制效率达90.75%;（2）3种用量的产紫青霉对烟草根结线虫的防治效果及烤烟综合农艺性状、产质量和均价均优于33.00 kg/hm2的淡紫拟青霉,其中以41.25 kg/hm2产紫青霉的防治效果、综合农艺性状、产值量和均价最好,平均防效为80.60%,烟叶产量、产值和均价分别为2119.50 kg/hm2、56762.77元/hm2和26.78元/kg。因此,产紫青霉K1在烟草根结线虫生物防治领域具有较大的潜能,可作为攀枝花烟草根结线虫病生防菌加以开发应用,推荐使用方法及用量为在烟草移栽时使用41.25 kg/hm2产紫青霉K1一次。 相似文献
11.
为探索AM真菌在牡丹组培上的应用效益及机理,保存和扩繁新优杂交牡丹资源及提供解决牡丹组培中生根难、移栽成活率低等瓶颈问题的新途径,选用牡丹新优杂交系种子进行胚培养得到胚根苗无菌体系,在瓶内对胚根苗接种AM真菌获得菌根化组培苗,继而研究AM真菌对牡丹组培苗生长发育和矿质元素吸收的影响.结果 表明,AM真菌能显著促进牡丹组... 相似文献
12.
为筛选能有效抑制常见植物病害的生防真菌,从甘肃省景泰县马铃薯连作田植株根际土壤中分离获得一株木霉菌株TS-1。通过形态学特征和EF-1α序列分析,将其鉴定为长枝木霉Trichoderma longibrachiatum。采用对峙培养法测定TS-1对6种病原菌的抑制作用,并利用该菌株不同浓度的分生孢子悬浮液处理小麦种子,分析种子形态学指标的变化。以产孢量为指标,采用单因素实验进行发酵条件的初探。结果表明：长枝木霉菌株TS-1对茄病镰孢(Fusarium solani)、接骨木镰孢(F. sambucinum)和灰葡萄孢(Botrytis cinerea)的抑菌率均在70%以上,抑菌效果较好。与对照相比,该菌株对小麦种子有一定的促生作用。通过对该菌株固体发酵条件初探,发现发酵培养基主要成分秸秆和麸皮的最佳配比为9:1、大麦添加量10%、碳氮源分别为乳糖和硫酸铵时产孢量最大。上述试验证明,长枝木霉TS-1具有较好的生防应用潜力,可进一步扩大培养。 相似文献
13.
CRISPR/Cas9系统是一种广泛应用于细菌、酵母、动物和植物中的基因组定点编辑技术,但该编辑系统的使用范围受PAM (proto-spacer-motif)位点NGG的限制。本研究通过突变Streptococcus pyogenes Cas9 (SpCas9)编码氨基酸(1135位的天冬氨酸D突变成缬氨酸V, 1335位的精氨酸R突变为谷胱氨酸Q, 1337位的苏氨酸T突变为精氨酸R,命名该突变子为Cas9-VQR)改造其识别PAM为NGA的位点以扩大其使用范围。并使用玉米Ubi启动子启动Cas9-VQR基因、优化SpCas9的密码子、加入保守的核定位信号序列、增加单子叶植物中保守的3′UTR序列和使用水稻U6启动子启动gRNA来修饰该编辑系统。结果表明Cas9-VQR系统能够识别PAM为NGA的位点,并进行有效的切割。体外酶切活性检测结果表明Cas9-VQR的切割效率为5%～70%。水稻转化检测结果表明Cas9-VQR的切割效率约为27.5%～70.5%,平均切割效率为46.23%。本研究拓宽了CRISPR/Cas9系统在作物中的使用范围,特别是NGA PAM位点较高的作物。 相似文献
14.
Teresa Paula Caams Immaculada Vias Josep Usall Carla Casals Cristina Solsona Neus Teixid 《Postharvest Biology and Technology》2008,49(1):86-95
The objective in the present study was to investigate the survival and effectiveness of different biological agent Pantoea agglomerans formulations against Penicillium spp. with different preharvest treatments. Results indicated a high sensitivity of non-adapted and osmotic-adapted P. agglomerans cells to environmental conditions in the field, resulting in preharvest treatments which were ineffective against Penicillium spp. In the second part of this study, dry conditions and solar radiation were identified as important environmental conditions that seriously affect populations of P. agglomerans cells. Different formulation strategies were tested in order to improve the resistance of cells to unfavourable environmental conditions. Osmotic-adapted P. agglomerans cells in the presence of 25 g L−1 of NaCl in the production medium [osmotic-adapted treatment (P25)] or at water activities (aw) of 0.98 [osmotic-adapted treatment (P98)] had higher survival rates than non-adapted cells, when these cells were sprayed on oranges and stored in hermetically sealed chambers at a low RH of 43%. Among seven additives tested, the presence of 5% Fungicover in the bacterial suspension improved adherence and persistence of P. agglomerans cells on oranges exposed to unfavourable conditions. Therefore, while P. agglomerans cells sprayed alone had log values of 0.5 CFU cm−2, in combination with Fungicover the population level of P. agglomerans cells reached log values of 5 and 4.2 CFU cm−2, at 0 and 24 h after application. Lyophilised cells showed greater resistance to unfavourable environmental conditions than fresh cells. The present study has demonstrated that the formulation improvement can provide better performance of biocontrol agents under environmental conditions non-conducive for growth and survival. 相似文献
15.
本研究旨在开发杀线虫活性高的生防资源以防控马铃薯腐烂茎线虫病。以34个细菌菌株发酵液为试材,通过直接触杀法测定其对马铃薯腐烂茎线虫的触杀活性;利用16S rDNA基因,并结合菌落形态和生理生化特征,确定生防菌株的分类地位。筛选出9个细菌菌株对马铃薯腐烂茎线虫的触杀活性较高,48 h线虫的校正死亡率达72.36%~82.59%;经鉴定,菌株Chi9-7、Nong3-5、Nong5-6和b1-3为枯草芽胞杆菌,菌株b5-4为贝莱斯芽胞杆菌,菌株Chi9-2、Xia4-11、Gu4-7和Z-1为解淀粉芽胞杆菌。本研究筛选并鉴定了杀线虫作用明显的9株芽胞杆菌,为微生物杀线剂的研发提供了新的生防资源,也为马铃薯腐烂茎线虫病的有效防控奠定了基础。 相似文献
16.
依据形态学、分子生物学以及菌株的生理生化特征对菌株A4进行鉴定,并研究其对樱桃番茄采后病害的防治效果。结果表明：菌株A4为生防菌贝莱斯芽孢杆菌（Bacillus velezensis）,其对樱桃番茄采后的重要致病菌灰霉菌（Botrytis cinerea）和扩展青霉菌（Penicillium expansum）均有抑制作用;平板对峙试验表明,贝莱斯芽孢杆菌A4对上述2种病原菌生长的抑制率分别高达66.56%和59.19%,预先接种浓度为1.0×109 CFU/mL A4菌悬液的樱桃番茄患病率分别降至14.67%和4.00%;扫描电镜显示,生防菌A4通过抑制病原菌的萌发和生长,降低病原菌的致病能力,减少病害的发生。本研究为生防菌贝莱斯芽孢杆菌的鉴定提供了参考,并为提高樱桃番茄采后病害的防治技术提供了新思路。 相似文献
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建立快速、准确、高通量与简便易行的基因型分析技术对基因功能解析、分子育种与突变体鉴定研究具有重要价值。本研究的目标是利用Cas9或Cas9NG变体与单分子指导RNA (single guide RNA, sgRNA)核糖核蛋白复合体(sgRNA/Cas9-RNP或sgRNA/Cas9NG-RNP)体外DNA定点内切酶活性,建立与优化简便高效与低成本的基因型分析技术。以我们前期创制的CRISPR/Cas9定点编辑玉米ZmWx基因第7外显子区域定点突变基因编辑后代分离群体为材料,以ZmWx靶位点两侧特异引物扩增的PCR产物为底物,利用原核表达并纯化的Cas9或Cas9-NG蛋白为DNA内切酶,以体外转录的靶向ZmWx基因靶点的sgRNA或骨架序列优化的sgRNA(enhancedsgRNA,esgRNA)为Cas9或Cas9-NG酶定点活性指导元件,通过体外组装为sgRNA/Cas9-RNP复合体,对目标样本迚行酶切,以区分目标位点野生型、纯合突变体、杂合突变体基因型,并对反应体系迚行了优化。研究表明,基于esgRNA/Cas9的PCR/RNP检测技术可对ZmWx基因编辑目标突变体后代迚行快速有效的基因型鉴定;实验体系优化结果表明, esgRNA/Cas9蛋白质量比为1∶1,各为1?g,20?L反应体系,37℃酶切0.5h,可对500ng待测DNA底物充分酶切并确定基因型;esgRNA/Cas9NG反应体系优化结果表明,当esgRNA与Cas9-NG蛋白均为2μg时, 37℃酶切4 h,可对500 ng DNA底物迚行酶切并实现基因型分析。利用Cas9NG拓宽靶位点检测范围的研究结果,暗示Cas9NG是以牺牲核酸酶酶切活性为代价降低了Cas9蛋白对PAM (protospacer adjacent motif, PAM)基序NGG序列的依赖性,实现PAM-NG基序识别能力, sgRNA/Cas9NG检测效率等仍有待提升与优化。本研究为基因功能解析、分子育种与突变体鉴定等研究提供了一套简便、成本低廉的技术方法,Cas9NG体外内切酶活性及其效率也为该Cas9突变体活体基因编辑技术研収提供了参考数据。 相似文献
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为探索丛枝菌根真菌(AMF)在牡丹组培中的应用,解决牡丹组培苗生根难、移栽成活率低等瓶颈问题,培育优质牡丹菌根化组培苗,以多个牡丹品种和杂交系组培苗为接种对象,设计筛选AMF接种剂种类和接种时期的最佳组合。结果表明：瓶内接种,在根长≥2.0 cm时接种AMF的菌根苗,AM菌根的侵染率与侵染强度最高,根长和根数分别比不接种AMF（对照）增长了49.29%和46.99%。AMF对不同品种的影响不一致,对于‘Z1-10-3’,接种混合孢子Glomus geosporum+Glomus mosseae(Gg+Gm)的侵染率(93.12%)与侵染强度(31.23)最高,极显著(P=0.01)高于单一接种剂Glomus mosseae(Gm)和Glomus geosporum(Gg);‘Z1-WM’接种Gg+Gm的侵染率(95.96%)与侵染强度(33.33)也最高,其次是Gg,然后是Gm;对于‘乌龙捧盛’,接种Gg的侵染率(85.76%)和侵染强度(25.80)最高。瓶内接种AMF能更有效地提高牡丹组培苗移栽成活率45.88个百分点。因此,牡丹组培苗在瓶内根长≥2.0 cm时接种混合AMF孢子,可实现组培苗菌根化过程,且其移栽成活率有效提高。 相似文献
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水稻潜根线虫在世界各稻区均普遍发生,给水稻生产造成一定的经济损失。从前期对根结线虫有较好毒杀活性的9个菌株中筛选出对水稻潜根线虫有很好毒杀效果的菌株。结果表明,2周发酵液2.5倍稀释液处理水稻潜根线虫,菌株ZW-1和XO-2对潜根线虫毒杀活性较强,72h的死亡率分别为93.7%和96.7%,LC50值分别为90.4451和76.3875μL/mL。根据形态学特征和rDNA-ITS序列分析,鉴定菌株ZW-1和XO-2分别为日本曲霉(Aspergillus japonicus)和黑曲霉(A.niger)。这些生防真菌菌株的发掘,为植物线虫病害的生物防治提供了新的微生物资源。 相似文献