二穗短柄草未成熟胚愈伤组织诱导及高频再生体系的建立

n以二倍体ABR 6和六倍体ABR 102二穗短柄草的幼胚为外植体,研究糖类、染色体组倍性和激素对愈伤组织诱导、分化及生根的影响。结果表明高效再生体系为愈伤组织诱导培养基：LS+2,4-D 2.5 mg•L-1+麦芽糖30 g•L-1+琼脂6.5g•L-1;愈伤组织继代培养基：LS+2,4-D 1.0 mg•L-1+6-BA0.2 mg•L-1+麦芽糖30g•L-1+琼脂6.5g•L-1;愈伤组织分化培养基：LS+KT 0.2mg•L-1+6-BA 0.5 mg•L-1 + CuSO4 0.6 mg•L-1+麦芽糖30g•L-1+琼脂6.5g•L-1;愈伤组织生根培养基：1/2 LS+IAA 0.6 mg•L-1 +麦芽糖20 g•L-1+琼脂7.0 g•L-1。优化了二穗短柄草愈伤组织培养条件,在含有30g•L-1麦芽糖的培养基上,愈伤组织出愈率最高可达96.67%,在含有0.2mg•L-1KT的培养基上,分化率最高为71.25%,进一步研究了影响短柄草再生的主要因素。     

农杆菌介导短柄草遗传转化体系的建立

为了探索短柄草的遗传转化体系,以二倍体短柄草ABR 6为受体材料,通过对诱导培养基类型、潮霉素筛选浓度和根癌农杆菌侵染浓度等参数的优化,建立了农杆菌介导短柄草遗传转化体系。结果表明,来源于未成熟胚的胚性愈伤组织在LS培养基上诱导率最高,达76.27%,最佳Hpt筛选浓度为40 mg·L^-1,最佳农杆菌侵染浓度为OD₆₀₀=0.6,在此条件下ABR 6的转化效率可达5%;通过PCR检测12株抗性植株,发现7株能扩增出Hpt基因（845 bp）条带;通过荧光显微镜观察转基因植株叶片,发现绿色荧光蛋白的表达,进一步证实了转基因植株的可靠性。     

氨基酸对小麦成熟胚愈伤组织诱导率的影响

为优化小麦成熟胚愈伤组织诱导体系,以中国春（Chinese Spring）成熟种子为材料,MS培养基为基础培养基,研究酒精、双氧水不同的灭菌时间和不同氨基酸组分对小麦成熟胚愈伤组织诱导与分化的影响。结果表明,当75%酒精浸泡时间控制在180 s、30%双氧水灭菌时间15 s以上时和当30%双氧水消毒时间控制在180 s、75%酒精浸泡时间15 s以上时,成熟胚均不会出现污染。甘氨酸、天冬氨酸、脯氨酸和亮氨酸是小麦成熟胚诱导的必须氨基酸。正交实验结果表明,培养基中最佳的氨基酸配组为甘氨酸60.0 mg·L^-1、天冬氨酸2.0 mg·L^-1、脯氨酸2.0 mg·L^-1和亮氨酸0.1 mg·L^-1。利用该培养体系,小麦成熟胚的出愈率达到97%以上。本研究优化了小麦成熟胚的培养体系,也为研究小麦氨基酸代谢的生理过程提供了参考。     

2-脱氧葡萄糖作为小麦愈伤组织筛选剂的研究

传统的转基因育种所带来的食品安全和环境安全问题是研究者无法回避的,选用生物安全性标记基因对转基因愈伤组织进行筛选是解决这一问题的重要途径。为寻找一种新型的生物安全型筛选剂用于小麦愈伤组织的筛选,并确定其最佳使用浓度,尝试利用2-脱氧葡萄糖（2-DOG）作为小麦遗传转化过程中愈伤组织的筛选剂,设置了0、400、600、800、1 000和1 500 mg·L^-1六个浓度梯度对4种小麦基因型的愈伤组织的再生进行了筛选。结果表明,800 mg·L^-1的2-DOG浓度为合适的筛选浓度,筛选时间以3周较合适。     

二穗短柄草成熟胚再生体系建立及农杆菌介导转化研究

为建立二穗短柄草组织培养及遗传转化体系,以二穗短柄草BD21-3成熟胚为外植体,对成熟胚愈伤诱导、分化以及农杆菌侵染条件进行了研究。结果表明,在含有2.5mg·L~(-1) 2,4-D的培养基上,愈伤组织出愈率最高为93.83%;在含有0.2mg·L~(-1) KT的分化培养基上,分化率最高为38.18%;对二穗短柄草胚性愈伤组织农杆菌转化和GUS染色结果表明,侵染的过程中农杆菌菌液浓度OD_(600)为0.6、侵染时间为5min时转化率最高。     

愈伤组织再生马铃薯脱毒苗生产体系的建立

以马铃薯茎尖、茎段、全叶、半叶为外植体,进行愈伤组织的诱导、继代、根芽分化和植株再生的研究,建立马铃薯脱毒再生体系。结果表明:(1)马铃薯茎尖、茎段、半叶、全叶在MS+6-BA1.5 mg·L-1+NAA1 mg·L-1+3%蔗糖+0.6%琼脂培养基中,愈伤组织诱导率分别达到97.3%、75.6%、96.8%、97.4%;(2)在MS+6-BA1.5mg·L-(1或ZT2.0 mg·L-1)+NAA0.5 mg·L-1+GA3 5 mg·L-1+3%蔗糖+0.6%琼脂培养基中不同外植体芽诱导率在15.4%～67.7%,根的分化率21.3%～88.2%。     

重金属Pb2+、Cd2+胁迫对青稞幼苗抗氧化能力的影响

为了解重金属对青稞幼苗的伤害机理以及青稞对重金属胁迫的抗性作用,以青稞幼苗为实验材料,采用水培方法研究了50 mg·L^-1 Pb²⁺、Cd²⁺胁迫对青稞幼苗叶片相对含水量（RWC）、丙二醛（MDA）和脯氨酸含量、抗氧化酶活性的影响。结果表明,50 mg·L^-1 Pb²⁺ 胁迫下青稞幼苗的叶片相对含水量（RWC）变化不大,丙二醛（MDA）和脯氨酸含量、过氧化物酶（POD）和过氧化氢酶（CAT）活性在胁迫后期有所升高,超氧化物歧化酶（SOD）活性在前期升高比较明显,其后缓慢下降。50 mg·L^-1 Cd²⁺胁迫下青稞幼苗叶片RWC明显减少,MDA含量显著增加,脯氨酸含量先急剧升高后下降,POD、CAT活性随着胁迫时间的增加而显著增强,其中CAT活性在胁迫末期稍有下降,SOD活性先明显升高后下降。综合来看,青稞幼苗在受到相同浓度Pb²⁺、Cd²⁺胁迫时,对Pb²⁺的抵抗能力大于Cd²⁺。     

几种生物制剂对小麦根腐病菌的毒力

为探索生物制剂对小麦根腐病的防控效果,采用菌丝生长法测定了6种生物制剂对禾谷镰刀菌、刺腐霉菌和小麦离蠕孢菌的毒力。结果表明,申嗪霉素、乙蒜素和春雷霉素具有良好的抑菌效果,其中申嗪霉素的毒力最强,对禾谷镰刀菌、刺腐霉菌和小麦离蠕孢菌的EC₅₀值分别为0.350 2 mg·L^-1、0.864 5 mg·L^-1和0.134 1 mg·L^-1,乙蒜素对刺腐霉菌和小麦离蠕孢菌的EC₅₀值分别为2.957 3 mg·L^-1和2.342 7 mg·L^-1,春雷霉素对刺腐霉菌和禾谷镰刀菌的EC₅₀值分别为0.864 5 mg·L^-1和5.090 9 mg·L^-1。室内盆栽试验表明,申嗪霉素和乙蒜素对禾谷镰刀菌的防治效果显著优于春雷霉素,防治效果分别为75.73%和74.68%;乙蒜素对刺腐霉菌的防治效果最好,防效为86.28%;申嗪霉素和乙蒜素对离蠕孢菌的防效分别为87.68%和83.25%。综上所述,申嗪霉素和乙蒜素能有效抑制禾谷镰刀菌、刺腐霉菌和小麦离蠕孢菌三种病原菌的生长,对小麦根腐病防治具有很大应用潜力。     

喷施硒肥对黑小麦籽粒产量及硒含量的影响

为探究叶面喷施硒肥对黑小麦籽粒产量和硒含量的效应,以临黑131和冬黑1206两个黑小麦品种为材料,分别在拔节期（4月17日）、孕穗期（4月28日）和灌浆期（5月14日）3个时期喷施0、10、20、30 mg·L^-14个浓度的亚硒酸钠溶液,分析叶面喷施不同浓度硒对黑小麦籽粒产量及硒含量的影响。结果表明,拔节期和孕穗期喷施硒肥对黑小麦有增产效果,而灌浆期喷施硒肥增产效果不明显;孕穗期和灌浆期喷施硒肥浓度为10 mg·L^-1至20 mg·L^-1时,黑小麦籽粒可达到富硒标准（0.15～0.30 mg·kg^-1）。综上所述,在黑小麦孕穗期叶面喷施浓度介于10 mg·L^-1至20 mg·L^-1之间的Na₂Se0₃溶液,在不减少产量的前提下,可显著提高黑小麦籽粒中的硒含量,使黑小麦籽粒达到富硒水平,增加黑小麦的营养价值。     

小麦质膜Na+/H+逆转运蛋白TaSOS1的表达分析

为探讨小麦质膜Na⁺/H⁺逆转运蛋白TaSOS1在逆境条件下的生物学功能,采用荧光定量PCR（RTPCR）方法,对 TaSOS1基因在4种不同胁迫条件下（200 mmol·L^-1 NaCl、4℃低温、100 μmol·L^-1 ABA和15% PEG6000）的表达水平进行了定量分析。结果表明,在正常条件下,小麦根中 TaSOS1的mRNA表达量要高于叶中的表达量。盐胁迫处理下,根中 TaSOS1特异性上调表达,说明 TaSOS1基因在根中的功能更强,其表达与盐胁迫有关且具有组织特异性;在4℃低温处理下, TaSOS1在叶中上调表达;而在ABA和PEG6000两种处理条件下, TaSOS1的表达没有规律性,推测 TaSOS1的表达途径可能为非ABA依赖型且与盐离子的特异性有关。生物信息学分析的结果表明,小麦质膜Na⁺/H⁺逆转运蛋白TaSOS1与拟南芥和水稻的质膜Na⁺/H⁺逆转运蛋白AtSOS1和OsSOS1具有高度的同源性,由此可以推测小麦TaSOS1具有与AtSOS1和OsSOS1类似的功能,可以把植物体内Na⁺排出体外,以减轻Na⁺对植物的毒害。     

Extreme resistance to infection by potato virus X in genotypes of wild tuber-bearingSolanum species

Summary Clones derived from thirty-one different accessions (nineteen of Argentine origin) belonging to eightSolanum species were screened for resistance to infection by potato virus X strain cp (PVX cp) by mechanical inoculation of plantlets that had been micropropagated in vitro. Estimates of PVX multiplication obtained by enzyme linked immunosorbent assay and slot blot nucleic acid hybridization allowed the identification of resistant clones derived from five accessions belonging toS. commersonii S. oplocense, S. sparsipilum andS. tuberosum andigena. Resistant genotypes supported PVX concentrations 5 to 15 times smaller than did the susceptible control cultivar Spunta. Graft inoculation test confirmed the presence of extreme resistance similar to that conferred by the ‘immunity’ gene X¹ (also called RX_act).     

Mineral composition of some traditional Mexican teas

Teas of plant origin traditionally consumed by the Mountain Pima of Chihuahua, Mexico, were analyzed for mineral nutritional content. Fe, Cu, Zn, Ca, and Mg composition was determined for native teas made from shoots ofTagetes lucida, T. filifolia, Elytraria imbricata, andHolodiscus dumosus, and from root xylem ofCeanothus depressus andPhaseolus ritensis. Native uses of these teas are also described.     

Effect of a continuous hot water treatment of potato tubers on seed-borne fungal pathogens

Summary The viability of five pathogens was decreased by treatment with hot water when tested in vitro.Polyscytalum pustulans was most sensitive andRhizoctonia solani least sensitive. Potato tubers were exposed to 55°C for 5 min in a commercial continuous hot water treatment plant using naturally contaminated seed tubers and tubers which had been inoculated by dipping in comminuted cultures. The frequency of eyes colonised byP. pustulans, Helminthosporium solani, andR. solani was reduced to virtually zero and the effect persisted on tubers subsequently stored at 4°C and at 15°C for up to 16 wk. Results withColletotrichum coccodes were inconclusive. Treatment suppressedPenicillium spp. which, however, rapidly recolonised the eyes during storage, leading to higher contamination levels in the treated than in the untreated tubers. With tubers inoculated withPhoma foveata, good control was achieved when the incubation period before treatment was 10 d but not when the period was 42 d.     

Field assessment of the relative importance of different aphid species in the transmission of potato virus Y

Summary Flying aphids were trapped throughout the summer on a vertical net downwind of a plot of PVY-infected potato plants. Of 6769 individuals caught, 165 transmitted PVY to tobacco test seedlings. Of 119 species or species groups caught, 20 were found to be vectors of which nine had not been recorded previously.Brachycaudus helichrysi, Myzus persicae, Phorodon humuli andAphis species accounted for 90% of transmissions andB. helichrysi alone for 52% of transmissions. The prospects of using this information to assess the amount of virus spread in a potato crop and of forecasting the timing and abundance of the main vectors are discussed.

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Zusammenfassung Im Abwind einer Parzelle mit PVY-infizierten Kartoffelpflanzen (7% PVY^O, 4% PVY^N, 89% beide) wurden fliegende Blattl?use in zwei vertikalen Netzen (4,4 m × 1,4 m mit hexagonalen L?chern mit 1,5 mm Durchmesser) gefangen und anschliessend auf Tabaks?mlinge überführt, um zu prüfen, ob sie PVY übertragen. Die Blattl?use wurden vom 8. Juni bis zum 8. August 1984 normalerweise an einem Tag pro Woche von 10 bis 17 Uhr gefangen. Im Laufe von 12 Fangtagen wurden 6769 sachte 52% der übertragungen bei nur 15% der totalen Fangzahl (Tab. 3).B. helichrysi mitMyzus persicae, Phorodon humuli und Species des GenusAphis ergaben sich als 90% der übertragenden Individuen bei nur 32% der gefangenen Proben. Acht Prozent der gefangenenB. helichrysi waren übertr?ger, dagegen nur 4,7% vonM. persicae. B. helichrysi k?nnte als Vektor besonders wichtig sein, weil sie in der Vegetationszeit Blattl?use von 119 Species oder Speciesgruppen gefangen, von denen 165 (2,44%) Tabak mit PVY inokulierten (Tab. 1). 133 Blattl?use übertrugen PVY^O und 32 übertrugen PVY^N (und m?glicherweise auch PVY^O). 20 Species oder Speciesgruppen waren Vektoren, von denen 9 (Cryptomyzus ballotae, Hyperomyzus lactucae, Metopolophium festucae, Myzus ligustri, M. myosotidis, Myzaphis rosarum, Sitobion avenae, S. fragariae undUroleucon spp.) erstmalig beschriebene Vektoren sind.Brachycaudus helichrysi verurfrüh auftritt (Tab. 2), wenn die Pflanzen am anf?lligsten sind und die Virus-Translokation zu den Knollen am schnellsten stattfindet. Die Aussichten zum Gebrauch dieser Information zur Absch?tzung des Aufbaues von Virusbefall im Bestand und die Aussichten für eine Vorhersage der Menge der wichtigsten Vektoren werden diskutiert. Es ist dringend erforderlich, die Biologie vonB. helichrysi zu untersuchen.

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Résumé Deux filets verticaux (4,40 m × 1,40 m à mailles héxagonales de 1,5 mm de diamètre) sont placés face au vent pour la capture des pucerons dans une parcelle de plants de pommes de terre contaminés par PVY (7% PVY^O, 4% PVY^N, 89% par les deux) puis les pucerons sont transferrés sur des plantules de tabac en milieu clos pour tester la transmission de PVY. Les captures de pucerons ont eu lieu du 8 juin au 8 ao?t 1984, en général à raison d'un journée par semaine, de 10 heures à 17 heures. Pendant les 12 premiers jours, 6769 pucerons de 119 espèces ou groupes d'espèces ont été capturés, les plantules de tabac ayant été inoculées par PVY avec 165 pucerons (2,44%) (tabl. 1). 133 pucerons étaient vecteurs de PVY^O et 32 de PVY^N (et probablement PVY^O). 20 espèces ou groupes d'espèces étaient des vecteurs dont 9 récemment enregistrés (Cryptomyzus ballotae, Hyperomyzus lactucae, Metopolophium festucae, Myzus ligustri, M. myosotidis, Myzaphis rosarum, Sitobion avenae, S. fragariae etUroleucon spp.).Brachycaudus helichrysi représentait 52% des transmissions mais seulement 15% du total capturé (tabl. 3).B. helichrysi, Myzus persicae, Phorodon humuli et les espèces du gèneAphis représentaient 90% des transmissions et 32% des captures seulement. 8,0% deB. helichrysi capturé a transmis le virus contre, 4,7% pourM. persicae. B. helichrysi peut être particulièrement important comme vecteur dans la mesure ou il appara?t t?t en saison (tabl. 2) lorsque les plantes sont très sensibles et la translocation di virus aux tubercules la plus rapide. Les perspectives à partir de cette connaissance sont discutées en vue d'analyser l'évolution du virus dans une culture et les prévisions de l'abondance des plus importants vecteurs. Une plus grande connaissance de la biologie deB. helichrysi devient urgente.

     

In vitro antimicrobial activity of different accessions ofSolanum commersonii Dun. from Uruguay

Summary The in vitro antimicrobial activity of extracts from accessions ofSolanum commersonii Dun. collected in the south of Uruguay was investigated against five microorganisms including the pathogenRalstonia solanacearum. A total of 30 extracts corresponding to organic and aqueous extracts were studied. Interestingly, most of the positive results for growth inhibition were againstR. solanacearum. The extracts were also analyzed for the presence of glycoalkaloids and lectins. Six of the ten aqueous extracts showed lectin presence and a wide variation in the type and amounts of glycoalkaloids, was found. Results indicate that there is no clear relationship between the antimicrobial activity against the five microorganisms screened and the presence or amounts of lectins and glycoalkaloids, traditionally regarded as possible antimicrobial metabolites in theSolanum genus, which indicates the presence of as yet unidentified antimicrobial compounds.     

玉米黑粉菌菌瘤内内生细菌对黑粉菌的冬孢子萌发的影响

利用高通量测序分析明确玉米黑粉菌菌瘤内菌粉培养液中内生细菌种类，结合16SrDNA分析，分离并鉴定在培养液中占比最高的菌株。以溶菌酶及Amp处理玉米黑粉菌冬孢子为对照，显微镜下观察菌瘤内内生细菌对玉米黑粉菌冬孢子萌发的影响。结果表明，在4份样品的菌粉培养液中，肠杆菌属细菌的占比在90%以上。当肠杆菌大量增殖时对黑粉菌的冬孢子萌发产生影响，可引起黑粉菌的冬孢子发生质壁分离，释放原生质体，部分原生质体再生成菌体；也可以使冬孢子壁直接被降解，原生质体萎缩。研究发现，玉米黑粉菌菌瘤内的内生细菌以肠杆菌为主，该内生细菌对黑粉菌的冬孢子萌发产生影响。     

Effect of haulm treatment and harvest time on incidence of tuber rots of potato (Solanum tuberosum L.) after standard wounding and on frequency of stem lesions caused byPhoma foveata Foister

Summary Various haulm treatments and harvest times differentially influenced the incidence of storage rots in potato tubers (Solanum tuberosum L.) in field experiments in Sweden performed over 4 years. The incidence of gangrene caused byPhoma foveata Foister (Phoma exigua var.foveata (Foister) Boerema) increased in cv. Bellona after spraying haulms with diquat as did the amount of infection caused byFusarium avenaceum (Corda ex. Fr.) Sacc. in all three cultivars (Bintje, Bellona, and Magnum Bonum). The lowestP. foveata infection levels occurred after pulling haulms. Late harvesting increased the frequency ofP. foveata in cvs Bintje and Bellona both at 2 and 4 wks interval between haulm treatment and harvest. Tuber and visible stem infection caused byP. foveata were correlated for cvs Bintje and Bellona, but not for cv. Magnum Bonum. The possible relation between tuber and stem infection is discussed.     

先玉335、郑单958和京科968抗虫能力的比较研究

以先玉335、郑单958、京科968为材料,以玉米螟(Ostrinia furnacalis)、黏虫(Mythimna separata)和禾谷缢管蚜(Rhopalosiphum padi)为供试昆虫,探究玉米品系大喇叭口期对玉米螟和黏虫及抽雄期对禾谷缢管蚜抗性差异,测定其主要抗虫化合物丁布类物质的含量。结果发现,京科968对玉米螟及禾谷缢管蚜抗性均高于另外两个品系;对黏虫的抗性,郑单958和京科968高于先玉335。大喇叭口期京科968中丁布类物质含量最高,抽雄期郑单958和京科968中主要丁布成分含量均高于先玉335,上述组织中丁布合成相关基因在郑单958及京科968中的表达量也均高于先玉335。本研究揭示杂交玉米抗虫性与抗性相关化合物含量及其相关基因表达间存在正向相关关系,为抗虫玉米分子育种提供理论参考。     

The effect of the moisture level of potting compost on the susceptibility of glasshouse-grown tubers to late blight

Summary The effect of the moisture level of potting compost at harvest on the susceptibility of tubers of three cultivars to late blight was investigated in the glasshouse in three successive years. Three moisture regimes (dry, moist, wet) were imposed at flowering time, the tubers harvested 3 weeks later and immediately dipped in a zoospore suspension ofPhytophthora infestans. Tubers from dry compost were significantly more susceptible than those from wet or moist compost, which showed similar levels of susceptibility.     

Evaluation of fungicides to control Petri disease pathogens in the grapevine propagation process

The efficacy of fourteen selected fungicides against Phaeomoniella chlamydospora and Phaeoacremonium aleophilum was evaluated in vitro by mycelial growth and conidial germination assays. Azoxystrobin, carbendazim and tebuconazole were the most effective fungicides against P. chlamydospora, while carbendazim and didecyldimethylammonium chloride were the most effective against P. aleophilum. An assay was also conducted with cubiet, hydroxyquinoline sulphate and didecyldimethylammonium chloride to determine their potential to prevent infections caused by these pathogens during the hydration stage in grapevine propagation. The results of this experiment demonstrated that P. chlamydospora and P. aleophilum were able to infect healthy cuttings during the hydration stage and showed that didecyldimethylammonium chloride was consistently the best treatment as a disinfectant soak for grapevine cuttings. Additionally, semi-commercial trials were conducted in two grapevine nurseries to evaluate the effectiveness of several strategies with carbendazim, flusilazole and hydroxyquinoline sulphate at different nursery stages. Soaking planting material in carbendazim during the hydration stage was the most effective treatment since neither P. chlamydospora nor P. aleophilum were isolated from planting material in either nursery.