不同烤烟品种青枯病的发生及农艺性状比较

为了筛选出对青枯病抗病性较好的高产烤烟品种，以黔抗1号、黔抗2号、黔抗3号、云烟119、K326、CF223、OX2028、NC196、PVH2254、金海1号、PVH1452等为供试烤烟品种，进行田间小区试验。通过调查、分析不同品种烟草青枯病的发生情况以及烟草农艺性状指标，评价不同品种间的差异。结果表明，以PVH2254品种青枯病发病最低，末次调查，其发病率及病情指数分别为11.3%和1.58，其次为PVH1452、NC196和黔抗2号等3个品种，青枯病发病较轻。在农艺性状上，以黔抗1号、黔抗3号表现最佳。建议在青枯病发生严重的植烟区，可考虑种植PVH 2254、黔抗2号、OX 2028或NC196品种，再辅以适当的营养调控和农业防治措施，以利于烟叶的可持续生产。     

表达Harpin蛋白质Hpa1功能片段的转基因小麦对赤霉病的抗性

来自水稻黄单胞菌的harpin蛋白质Hpa1有促进植物生长、诱导植物抗病性的功能，Hpa1序列10~42氨基酸片段（Hpa1_10-42）的活性比全长分子高1.3~7.5倍。为研究Hpa1_10-42在转基因小麦体内表达以后对赤霉病的影响和评价转基因小麦抗病水平与应用潜力，对6个小麦转基因系进行了测定。结果显示，Hpa1_10-42在转基因系T₃~T₅代呈现稳定表达，用禾谷镰刀菌一个分离物进行接种以后，转基因系发生赤霉病的程度较非转基因小麦显著降低，且转基因系T₃~T₅代赤霉病降低的趋势一致。另外，转基因系小麦病害减轻的程度，与在非转基因小麦上使用杀菌剂的效果相当，表明Hpa1_10-42转基因表达对小麦赤霉病有防治作用。     

低温胁迫下转ICE1基因水稻养分变化的研究

采用盆栽试验,选用转ICE1基因水稻T_4-8株系和T_4-9株系及未转基因水稻（Oryza sativa L.）品种垦鉴稻10号为试验材料,研究苗期低温胁迫对转基因水稻氮、磷、钾养分吸收的影响。结果表明,低温胁迫期间,转基因水稻幼苗丙二醛含量维持在较低水平,SOD活性维持在较高水平;地上部分氮、磷、钾含量变化不大,一直维持在较高水平;地下部分氮、磷、钾含量呈缓慢下降趋势,但降幅比非转基因水稻小,在胁迫第10天,T_4-8株系和T_4-9株系氮含量均高出对照13.86%,磷含量分别高出42.31%和50.00%,钾含量均高出85.71%。说明在低温胁迫条件下,ICE1基因过量表达可以增强水稻的抗寒性,减轻低温对水稻的伤害,保持根系较强的吸收养分能力。     

转ATCIPK23基因烟草植株钾素营养效率研究

通过考察转ATCIPK23基因烟草植株在不同钾浓度下钾及干物质积累的能力，研究了外源基因的导入对转基因烟草植株钾吸收能力的影响。结果表明，在低钾环境下，转基因株系具有明显的生长优势，烟株发育较快、根系发达、生物量较大、植株的含钾率也有明显提高，随着供钾量的减少，这种差异幅度逐渐增大。当供钾浓度为0.1 mmol·L^-1时，对照品种K326植株的平均钾含量为17.7 g·kg^-1，而转基因品系KA12为23.3 g·kg^-1，KA16为24.6 g·kg^-1，KA21为22.1 g·kg^-1，分别较对照品种增加了31.6%、39.0%和27.7%。钾吸收动力学研究表明，转基因植株与对照品种植株的最大吸收效率（V_max）差别不大，而最低浓度（C_min）与米氏常数（K_M）的差别较为明显。     

不同品种(系)烟草叶片对注射接种烟草青枯病菌的反应

烟草细菌性青枯病菌(Pseudomonas solanacearum)注射接种烟草叶片,不同烟草品种对青枯菌不同菌株及对同一菌株不同浓度的接种反应具有明显差异。单育3号、金星6007和永定一号经注射接种后症状较重,而005-1和K326的症状较轻。接种不同浓度的青枯菌,烟草叶片的症状发展规律相似,但随接种浓度的增高,症状加重。相同条件下,离体叶片症状表现较重。菌株4-11引起的症状最重,其次为菌株1-1,菌株3-5引起的症状最轻。     

温度对小麦矮腥黑穗病菌冬孢子萌发的影响

温度对小麦矮腥黑穗病菌冬孢子萌发影响的试验结果表明,TCK冬孢子在-2～12℃范围内都可以萌发,5℃为最佳萌发温度。同一分离菌冬孢子在不同温度处理的萌发特性和萌发率有差异;不同分离菌冬孢子在相同温度下的萌发特性和萌发率有很大差异。在5℃下5个分离菌中,冬孢子萌发起始时间最短的为15 d,最长为35 d,培养60 d时分离菌T_t1、T_t2、T_y、T_m和T_u冬孢子的萌发率分别为86.4%、23.9%、23.3%、44.3%和81.0%。根据分离菌T_t2不同温度下培养50 d时的萌发率,建立了萌发率(Y)与温度(X)的模型为Y=0.150 8 exp[-0.02949(X-4.957 6)]2,此结果为TCK的风险评估提供了基础数据。     

不同茬口对高寒阴湿区当归养分吸收特性和药材质量的影响

为建立高效经济的当归生产模式，于2017—2019年在甘肃省渭源县设置5种作物茬口（T₃：豌豆-小麦-当归，T₂：豌豆-蒙古黄芪-当归，T₃：豌豆-马铃薯-当归，T₄：豌豆-休耕-当归，CK：豌豆-当归-当归）进行田间试验，秋季收获后测定不同茬口当归根、叶养分含量、根活性成分和产量，采用主成分分析法确定各指标权重值、隶属综合因子分析法确定5种茬口的综合评价指数。结果表明：T₂处理有利于促进当归根、叶中氮、磷、钾吸收累积，根部氮、磷、钾含量较CK分别显著增加29.98%、32.96%、24.29%（P<0.05）、叶部氮、磷、钾含量较CK分别显著增加37.72%、42.08%、64.63%（P<0.05），且该茬口的当归产量、根的阿魏酸、浸出物含量较CK分别显著增加19.44%、17.37%、9.54%（P<0.05）。综合评价指数表现为T₂(0.74)>T₄(0.51)>T₃(0.48)>CK(0.29)>T₃(0.25)。综上可知，豌豆-蒙古黄芪-当归是高寒阴湿区当归高效经济生产的适宜模式。     

小偃9366抗条锈病基因的遗传分析及分子作图

为了明确小偃9366抗条锈病遗传特点,对小偃9366与铭贤169及其杂交F₁、F₂、F₃和BC₁F₁代进行温室苗期抗条锈性遗传分析,选取400余对SSR引物对接种CYR31的群体进行分子标记,并利用目标基因的侧翼引物分析99个黄淮麦区主栽小麦品种。小偃9366对CYR25的抗病性由1显、1隐2对基因独立控制,对CYR27的抗病性由3对显性基因控制,其中2对基因表现累加作用,对CYR30 和CYR31的抗病性均由1对显性基因独立控制,对Su11-4的抗性由2对隐性基因独立控制。位于2AL上的6个标记Xwmc794、Xwmc455、Xwmc261、Xgwm47、Xgwm294和Xcfd168与抗CYR31基因(暂命名Yrxy9366)连锁,与目的基因的遗传距离分别为10.8、6.5、3.2、4.4、16.0和32.8 cM,将Yrxy9366定位在2AL上。利用Xwmc261、Xgwm47两个引物分析99个黄淮麦区主栽小麦品种,仅5%检测到同源片段。研究表明Yrxy9366是一个新的抗病基因。     

烟草赤星病危害烤烟产量产值损失测定

在烤烟生长中后期 ,应用40%菌核净500倍液人为控制烟草赤星病发生程度 ,形成不同危害梯度 ,以研究烟草赤星病对产量产值的影响。经试验分析 ,建立数学模型。中上等烟产量损失率(Y₁)与病情指数(X)的关系式:Y₁=0.1516+1.2824X;产值损失率(Y₂)与病情指数(X)的关系式:Y₂=0.3239+0.9831X。     

毒株、苗龄及温度对烟草苗期接种TMV试验的影响

 病毒株系、苗龄、接种后培养条件是影响烟草病毒病发生发展的关键因素,为确保烟草接种TMV抗性鉴定和药效生测试验的准确性,在N基因烟草和普通烟草上接种TMV,采用病毒生物学、qRT-PCR和Western blot技术,研究病毒的分离纯化、株系、稀释限点、传导路径、病害症状,以及N基因抗TMV的温敏性。结果显示,TMV为U1株系,在K326上活体保存50~65 d,仍具有较强的致病力,稀释限点为10⁵,接种物浓度以10²稀释为宜。接种后TMV先由接种叶传导到主茎,向下至根、向上至心叶,然后是上部叶,最后传导至下部叶,12~16 d布满全株,21~28 d花叶畸形显著,病情调查宜在2~3周内。温度和苗龄影响N基因烟草对TMV的抗性反应,培养温度应在25℃~27℃,苗龄以6~7叶期为宜。     

酵母pac1基因介导对葡萄B病毒的抗性

为明确广谱性抗病毒基因—酵母pac1基因对葡萄B病毒(Grapevine virus B,GVB)的抗性效果,通过农杆菌介导的遗传转化,将pac1基因导入西方烟37B,对转基因植株进行PCR鉴定及Southern blot分析,通过病毒摩擦接种观察症状以及实时荧光定量RT-PCR检测植株体内病毒含量,并对转基因植株抗病性进行初步鉴定。结果表明,目的基因pac1成功导入并整合至西方烟37B基因组,共获得10个转基因株系。不同株系的T1代烟草中阳性植株比例为16.7%~72.7%,表明目的基因可成功遗传到子代。接种病毒后转基因植株普遍延迟发病,但后期症状与非转基因对照相似,其中仅1个转基因株系B6具有不表现典型症状等抗性反应。接种植株病毒含量检测中,所有转基因植株均检测到病毒存在,但表现为抗病的B6株系中病毒含量显著低于非转基因对照,表明该转基因植株虽不能完全抵抗GVB侵染,但对GVB具有耐病性。     

含双病原物诱导启动子无选择标记转基因烟草的获得

Transgenic plants are controversial for their edible and environmental security. Marker-free transgenic plants can be produced by the construction and transformation of plant expression vectors carrying twin T-DNAs. The construction of plant expression vectors harboring twin T-DNAs and two pathogen-inducible promoters was previously reported. These vector plasmids were introduced into tobacco plants and the transgenic tobacco plants were obtained. In this paper we report that T₁ transgenic tobacco seedlings were produced through classical genetics approach. Analysis of the seedlings demonstrated that some of them were marker-free lines. The segregation of exogenous genes in T₁ transgenic tobacco seedlings was tested by using two methods. Firstly, the ability of resistance to kanamycin was analyzed in T₁ transgenic tobacco seedlings from 14 transgenic plant lines. It was found that the segregation ratio of NPTII genes met well with Mendel’s law in 13 transgenic tobacco lines. So it was deduced that NPTII gene was as a single copy integrated into one of the homologous chromosomes. Then the NPTII genes and uidA genes of 130 T₁ transgenic seedlings were detected from the above 13 tobacco lines. The results showed that uidA genes were only detected in 20.77% of the seedlings, NPTII genes were solely detected in 22.31% of the seedlings, but both exogenous genes were in 53.85% of the seedlings. The segregation ratio of the two genes was consistent with the law of independent assortment (9∶3∶3∶1). These results suggested that the selective marker gene had no linkage with the reporter gene and they were segregated independently in the T₁ transgenic tobacco plants. This method, as compared with traditional backcross, is confirmed a more easy and rapid way to eliminate the antibiotic resistance gene used as a selective marker in transgenic plants.     

Coat protein-mediated transgenic resistance in tomato against a IB subgroup Cucumber mosaic virus strain

Transgenic tomato plants containing the coat protein (CP) gene of Cucumber mosaic virus (CMV) of subgroup IB were developed through Agrobacterium-mediated transformations. The progenies of transgenic plants showed the presence of transgene, its expression and translation of 26 KDa CP. The T₁ and T₂ generation plants were evaluated for resistance against challenge inoculations by a homologous strain of CMV. Visual observations of challenged transgenic plants categorized them into resistant, tolerant and susceptible as compared with untransformed control plants. Out of 33 plants of the T₁ generation, 36.3% showed resistance and remained symptomless throughout their life, 48.4% showed tolerance which developed delayed symptoms of mild mosaic, and 15.1% showed susceptibility to CMV which developed severe systemic mosaic and leaf distortion symptoms after 30?days of virus challenge. Out of 120 plants of the T₂ generation, 60% showed resistance, 26.6% were tolerant and only 13.3% were found susceptible to challenge inoculations of CMV. Resistant transgenic plants also showed less CP accumulation in systemic upper leaves as compared with challenged untransformed plants. In this study, CP of a CMV subgroup IB strain has demonstrated a significant level of resistance in transgenic tomato plants against the CMV strain. The strategy may be applied for better quality and productivity of tomato crops.     

A short‐length single chimeric transgene induces simultaneous silencing of Agrobacterium tumefaciens oncogenes and resistance to crown gall

Transformation with self‐complementary oncogene sequences was used to silence the Agrobacterium tumefaciens oncogenes ipt and iaaM. The silencing response was triggered by using a very short chimeric sequence where conserved fragments from both oncogenes were fused in one unique transgene. Most T₀ transgenic tobacco lines and T₁ seedlings evaluated in vitro had intermediate or very low susceptibility to A. tumefaciens as compared with the wildtype plants. A greenhouse evaluation of whole plants confirmed the lines that were resistant. Low levels of transgene hairpin RNA (hpRNA) coupled with small interfering RNA (siRNA) accumulation correlated with oncogene silencing and, therefore, resistance to crown gall. After infection with the oncogenic strain, much lower levels of the oncogenes’ mRNA were found in resistant lines than in wildtype plants. The frequency of resistant lines, with few or no symptoms, produced with the chimeric construct was similar to the highest reported efficiencies obtained by using sense and antisense whole oncogene sequences.     

Development of insect‐resistant transgenic rice with Cry1C*‐free endosperm

BACKGROUND: Yellow stem borer (Tryporyza incertulas Walker), striped stem borer (Chilo suppressalis Walker) and leaf folder (Cnaphalocrocis medinalis Guenec) are three lepidopteran pests that cause severe damage to rice in many areas of the world. In this study, novel insect‐resistant transgenic rice was developed in which Bt protein expression was nearly absent in the endosperm. The resistant gene, cry1C*, driven by the rice rbcS promoter (small subunit of ribulose‐1,5‐bisphosphate carboxylase/oxygenase), was introduced into Zhonghua 11 (Oryza sativa L. ssp. japonica) by Agrobacterium‐mediated transformation. RESULTS: A total of 83 independent transformants were obtained, 19 of which were characterised as single‐copy foreign gene insertion. After preliminary screening of the T₁ families of these 19 transformants in the field, six highly insect‐resistant homozygous lines were selected. These six homozygous transgenic lines were field tested for resistance to leaf folders and stem borers, and for their agronomic performance. The Cry1C* protein levels in leaves and endosperm were measured by ELISA. Subsequently, the elite transgenic line RJ5 was selected; this line not only possessed high resistance to leaf folders and stem borers, normal agronomic performance, but also Cry1C* expression was only 2.6 ng g^?1 in the endosperm. CONCLUSION: These results indicated that RJ5 has the potential for widespread utility in rice production. Copyright © 2009 Society of Chemical Industry     

Preliminary studies on resistance phenotypes to Turnip mosaic virus in Brassica napus and B. carinata from different continents and effects of temperature on their expression

Preliminary studies were undertaken to establish the occurrence of Turnip mosaic virus (TuMV) resistance phenotypes in 99 Brassica napus and 32 B. carinata accessions, breeding lines and cultivars mostly from Africa, Australia, the Indian sub-continent or China. An isolate of TuMV pathotype 8 (WA-Ap1) was used in most inoculations. The influence of temperature on expression of resistance phenotypes was determined. Those identified were O (extreme resistance), R_N (localised hypersensitivity), R (resistance to systemic movement without necrosis), +_N (systemic infection with some necrosis), + (susceptibility), and R_N/+ (systemic infection with necrosis limited to inoculated leaves). In the initial glasshouse evaluations with B. napus, 18 lines developed phenotype O, 42 R_N, two + (both from Australia), and 30 segregated for O and R_N, two for R and R_N, one for O and R, two for O, R and R_N, one for O, R_N and +_N, and one for R_N and +_N. Phenotype +_N only occurred in two lines from India. In the initial glasshouse evaluations with B. carinata, 14 lines developed O, two R_N or R, and one +; the 13 remaining lines segregated for phenotypes O and R (12), or R_N and R (1). In B. carinata, phenotype R_N only occurred in African lines, and phenotype + only in a line from Pakistan. The 18 B. napus and 14 B. carinata lines that developed phenotype O uniformly were re-evaluated at low (16 and/or 18 °C) and high (25 °C) temperatures in the glasshouse three or two times, respectively, and again under controlled environment room conditions at 16 and at 28 °C. While in most lines phenotype O was replaced by other phenotypes or segregated with other phenotypes at the higher temperatures, it occurred uniformly regardless of temperature in Chinese B. napus line (06-6-3777) and three African B. carinata lines (IP 117, ST 18 and ST 50). Graft inoculations confirmed extreme resistance in these four lines. Other promising lines which displayed phenotype O in >75 % of inoculated plants included for B. napus Chinese lines 06-p71-1 and 06-p74-4 and French line Cresor, and for B. carinata African lines Mbeya Green, ML-EM-1, ML-EM-7 and ML-EM-8, and Australian lines P 195923.3 and 30200533. Five promising lines segregated for phenotype R_N, B. napus Ding 110, Hyola 42, Fan 028, ZY 007 and Qu 1104. Chinese B. napus line Ding 110 developed phenotype R_N uniformly at high (25 and 28 °C) temperatures. Thus, in plants developing phenotypes R_N, R and +, low temperature retarded virus multiplication in inoculated leaves to below the level at which it can be identified using ELISA. No clear phenotypic differences attributable to TuMV pathotype were found in tests in which isolates from pathotypes 1, 7 and 8 were used. This highlights the need for future evaluations for TuMV resistance in Brassica germplasm to be done at higher temperatures. This study also highlights the need to identify additional genes responsible for the different resistance and susceptibility reactions found, especially in B. carinata. The four lines that developed phenotype O uniformly at all temperatures and also withstood graft-inoculation will be particularly valuable for developing new TuMV-resistant cultivars of oilseed and forage Brassicas.     

Agronomic performance of F1, F2 and F3 hybrids between weedy rice and transgenic glufosinate-resistant rice

BACKGROUND: Studies of hybrid fitness, of which agronomic performance may be an indicator, can help in evaluating the potential for introgression of a transgene from a transgenic crop to wild relatives. The objective of this study was to assess the agronomic performance of reciprocal hybrids between two transgenic glufosinate‐resistant rice lines, Y0003 and 99‐t, and two weedy rice accessions, WR1 and WR2, in the greenhouse. RESULTS: F₁ hybrids displayed heterosis in height, flag leaf area and number of spikelets per panicle. The agronomic performance of F1 between WR1 and Y0003 was not affected by crossing direction. The tiller and panicle numbers of F1 individuals were higher than their F2 counterparts. However, these traits did not change significantly from the F2 to the F3 generation or in hybrids with weedy rice as maternal or paternal plants. For all hybrids, the in vitro germination rates of fresh pollen were similar and significantly lower than those of their parents, seed sets were similar to or of lower value than those of weedy rice parents and seed shattering characteristics were partially suppressed, but the survival of hybrids over winter in the field was similar to that of weedy rice parents. All F1, F2 and F3 hybrids had similar composite agronomic performance to weedy rice parents. CONCLUSION: There was no significant decrease in the composite agronomic performance of any of the hybrids compared with weedy rice. This implies that gene flow from transgenic cultivated rice to weedy rice could occur under natural conditions. Copyright © 2011 Society of Chemical Industry     

Expression of human protoporphyrinogen oxidase in transgenic rice induces both a photodynamic response and oxyfluorfen resistance

A human protoporphyrinogen oxidase (Protox) coding sequence under the control of a ubiquitin promoter was introduced into rice to determine whether transgenic rice overexpressing the human Protox gene exhibits resistance against a peroxidizing herbicide. The transgenic rice lines (H3, H4, H5, H6, H9, and H10) transcribed the human Protox mRNA, whereas hybridizing RNA band was not detected in wild-type rice, indicating that the human Protox gene had been successfully transmitted into transgenic rice plants. The transgenic lines H9 and H10 showed growth retardation and light-dependent formation of necrotic lesions. Compared with wild-type rice plants, rice with a human Protox gene had increased Protox activity and content of the photosensitizer protoporphyrin IX, and reduced chlorophyll. The photosynthetic efficiency in lines H9 and H10, as indicated by F_v/F_m, was not different from that of wild type. The two transgenic lines had decreased levels of antheraxanthin, lutein, and β-carotene and similar levels of neoxanthin and violaxanthin as compared with wild-type plants. The staining activities of catalase, peroxidase, superoxide dismutase, and glutathione reductase were higher in transgenic lines than in wild type. Line H9 germinated in the presence of 20 μM oxyfluorfen, whereas 2 μM oxyfluorfen inhibited the germination of wild-type seeds. Thus, the transgenic rice plants exhibited enhanced resistance to oxyfluorfen.     

Coat protein-mediated resistance to isolates of barley yellow dwarf in oats and barley

Tissue cultures of GAF30/Park oats were biolistically co-transformed with constructs containing the coat protein (CP) genes of the P-PAV, MAV-PS1 or NY-RPV isolates of barley yellow dwarf virus (BYDV), together with a construct containing the bar gene for herbicide resistance and the uidA reporter gene. Transformed, herbicide-resistant tissue cultures were screened by PCR for the presence of the CP genes. Fertile regenerated plants were recovered from some CP-transformed tissue cultures. T₁ progeny of these plants were screened for resistance to the BYDV isolate corresponding to the introduced gene by inoculation with viruliferous aphids followed by ELISA tests. Variation in ELISA values for GAF30/Park control plants made interpretation of the data difficult, but oat plants resistant to each of the three isolates of BYDV (ELISA values less than 0.3; virus titers equivalent to less than 25% of infected controls) were identified in T₁ generations. Further testing of MAV-PS1 CP-transformed lines to the T₂ generation, NY-RPV CP-transformed lines to the T₃ generation and P-PAV CP-transformed lines to the T₄ generation identified further resistant plants. Similarly, immature embryos and calli of the barley cultivar Golden Promise were biolistically bombarded with constructs containing the CP gene of the P-PAV isolate of BYDV and the bar and uidA reporter genes, lines of self-fertile P-PAV CP-transformed barley plants were developed, and T₁plants were screened for resistance to P-PAV. Eight plants from six lines showed moderate to high levels of resistance to P-PAV that correlated with the presence of the CP gene. Plants giving low ELISA values were also found in other lines, even though the CP gene was not detected in these plants. Some T₂ plants derived from resistant parents that contained the CP gene were themselves highly resistant.