白叶枯病菌效应子XopN在拥有OsSWEET11同源基因的水稻品种上发挥毒性作用

【目的】黄单胞菌细胞外泌蛋白质(Xanthomonas outer proteins, Xops)是植物病原黄单胞菌高度保守的毒性效应子或毒性辅助组分,通过细菌Ⅲ型分泌系统分泌到细菌细胞外部,然后转入植物细胞而发挥病理作用。水稻黄单胞菌水稻致病变种即水稻白叶枯病菌的标准菌株PXO99^A分泌的XopN是一种毒性效应子,通过影响寄主免疫反应而使水稻发病。但是,XopN对病菌毒性的影响是否因水稻品种的不同而异,还有待研究。【方法】利用同源双交换技术,先敲除了PXO99^A的XopN基因,获得了∆XopN突变体,又经过遗传互补,得到了回补菌株∆XopN/XopN。通过营养肉汤液体培养,测定了XopN对病菌繁殖能力的影响;根据文献选用14个水稻品种,通过接种实验,测定了PXO99^A对这些品种的毒性与XopN敲除或回补的影响;在XopN发挥毒性作用的水稻品种上,测定了隐性抗病基因OsSWEET11/xa13与显性感病基因OsSWEET11/Xa13受病菌侵染而表达的情况,分析了XopN敲除或回补的影响。【结果】在营养肉汤液体培养过程中,病菌突变体菌株∆XopN的繁殖速度明显低于野生型PXO99^A。PXO99^A、∆XopN和∆XopN/XopN接种水稻后,根据其在水稻叶片组织内的繁殖量及随后产生的白叶枯病症状的严重程度,将供试的14个水稻品种分为两种情况。一是感病程度与病菌XopN是否敲除或回补无关,这有10个水稻品种（IRBB1、IRBB3、IRBB8、IRBB10、IRBB14、IR24、IRBB203、IRBB204、IRBB205和IRBB211）,PXO99^A、∆XopN和∆XopN/XopN对它们的毒性无明显差别。二是XopN对病菌毒性发挥作用的水稻品种,包括高度感病的3个品种（IRBB208、Asominori和日本晴）和低感品种IRBB13。与PXO99^A或∆XopN/XopN相比,∆XopN对这4个水稻品种的毒性大为降低。在IRBB13上,病菌侵染对隐性抗病基因OsSWEET11/xa13在叶片内的表达发生抑制作用,这一效应与XopN的毒性功能相关。相反,日本晴显性感病基因OsSWEET11/Xa13却受病菌侵染的诱导,在叶片内的表达水平大幅度提高。IRBB208和Asominori携带OsSWEET11/Xa13同源基因,该同源基因在叶片内的表达水平也因病菌侵染而大幅提高。在这4个水稻品种上,PXO99^A和∆XopN/XopN能够诱导OsSWEET11/Xa13或其同源基因表达,但∆XopN无此作用。另外,XopN对病菌在非寄主植物烟草上诱发过敏反应有量变贡献,相比PXO99^A或∆XopN/XopN,∆XopN引起过敏反应的程度有所降低。【结论】XopN是一个有限广谱性效应子,在拥有OsSWEET11同源基因的水稻品种上发挥毒性作用。XopN也是病菌繁殖所需要的,对病菌在非寄主植物上诱导过敏反应有一定贡献。     

Genome-wide association study for spot blotch resistance in Afghan wheat germplasm

Spot blotch (SB), caused by Bipolaris sorokiniana, is a devastating disease of wheat globally, especially in South Asia and South America. Understanding the genetics of resistance to SB is important for developing breeding strategies to improve resistance. A panel of 301 genotypes from Afghanistan was phenotyped over two crop seasons using a mixture of virulent B. sorokiniana isolates and genotyped using DArTSeq to obtain genome-wide markers. Fifty genotypes (16.6%) showed disease scores less than the resistant control. Principal component analysis using the genotypic data clustered the genotypes into five different groups. Among models used for genome-wide association mapping, the multilocus mixed model, and fixed and random model circulating probability unification algorithms were most effective in identifying significant marker-trait associations (MTA). Twenty-five MTAs at p ≤ .001 were identified on chromosomes 1A, 1B, 1D, 2B, 2D, 3A, 3B, 4A, 5A, 5B, 6A, 7A, and 7D, indicating the quantitative nature of resistance to SB. Phenotypic variation explained by these markers ranged from 2.0% to 17.7%, and genomic regions on the chromosomes 1D, 2D, 3A, 3B, 4A, 5A, and 5B coincided with loci identified in previous studies. Three single nucleotide polymorphism (SNP) markers on chromosomes 1B (SNP 1113207) and 5A (SNPs 5411867 and 998276) were significant in both crop seasons as well as in the combined analysis across seasons. Marker 5411867 is close to Vrn-A1, shown to be associated with SB in previous studies. Furthermore, among known SB resistance genes, Sb2 on chromosome 5B was predicted to be significant in this panel.     

Regional and temporal differentiation of virulence phenotypes of Puccinia triticina from common wheat in Russia during the period 2001–2018

Leaf rust, caused by the fungus Puccinia triticina, is one of the most damaging rust diseases of wheat in Russia. Populations of P. triticina were monitored in seven regions of Russia from 2001 to 2018, with a total of 5,191 single urediniospore isolates from bread wheat (Triticum aestivum) being analysed. Populations have changed significantly in all regions since 2012, after 2 years of drought (2010–2011). Regional collections of P. triticina were also significantly different between the two periods 2001–2009 and 2012–2018, with changes along two geographic gradients from West Siberia to the north-west and south-west (North Caucasia) of the European part of Russia. All tested isolates were avirulent to resistance gene Lr9 in 2001–2009 but, since 2010, virulence to Lr9 has occurred and annually increased in the Asian part of Russia (Ural and West Siberia) due to deployment of cultivars with the Lr9 gene. Virulence to Lr2a and Lr15 was considerably lower in Dagestan (6%–33%) and all European regions (35%–67%) than in Asian regions (84%–96%). During 2001–2009, virulence on Lr1 was also lower in Dagestan (33%) and the European regions (50%–77%) than in Asia (91%–96%); however, by 2012–2018, nearly all isolates were virulent on Lr1. Remarkable changes were observed in frequencies of P. triticina races defined by their virulence/avirulence to Lr1 and Lr2a genes. We postulate the P. triticina population in Dagestan is specific to that area and pathogen populations in European and Asian parts of Russia are distinct.     

Comparison of growth,survivorship, seed production and shedding of eight weed species in a wheat crop in Western Australia

Development of integrated weed management strategies is dependent on a thorough knowledge of the demography of individual species. The current research established eight winter or summer weed species in a winter annual wheat cropping system at Wongan Hills, Western Australia, and investigated emergence of the first cohort of each species, survivorship, plant size, seed production and seed shedding over three years (2016–2019). The winter weeds Bromus diandrus and Lolium rigidum emerged at the same time as the wheat crop, and the initial cohort of marked plants had 100% survival to seed production in each year. By comparison, other winter weed species like Hordeum leporinum, Rumex hypogaeus, Sonchus oleraceus and Polygonum aviculare frequently emerged later than the crop and had a lower percentage of plants surviving to seed production. However, individual S. oleraceus and P. aviculare plants had the greatest seed production compared to other species. All winter weeds had variable patterns of seed shedding between years, with the exception of L. rigidum. Summer weed species emerged at the same time, but plants in the initial cohort of each species did not always survive to produce seed. The early emergence and high survivorship of B. diandrus indicates high competitive ability, but shedding commenced at a similar time to L. rigidum and harvest weed seed control may be a viable control method for this species.     

Influence of intra-row cruciferous surrogate weed growth on crop yield in organic spring cereals

In Northern Europe, inter-row hoeing has become a popular tactic for controlling weeds in organic cereals. Hoeing is highly effective and can be implemented from crop emergence until stem elongation to maintain a nearly weed-free inter-row zone. However, hoeing has a lesser effect on weeds growing in the intra-row zone, where crop–weed proximity results in heightened competition. In the hoed cereal system, it is investigated whether tall-growing, competitive, cruciferous weeds in the intra-row zone affect crop biomass, yield and thousand kernel weight (TKW). An additive experimental design is employed to enable the fitting of rectangular hyperbolas, describing and quantifying the effects of increasing intra-row surrogate weed density on crop growth parameters. Regressions were studied under the influence of crop (spring barley and spring wheat), row spacing (narrow [12.5 or 15.0 cm] and wide [25.0 cm]) and nitrogen rate (50 and 100 kg NH₄-N/ha). Cruciferous surrogate weeds were found to impact crop yield and quality severely. For example, ten intra-row plants/m² of surrogate weed Sinapis alba reduced grains yields by 7%–14% in spring barley and by 7%–32% in spring wheat with yield losses becoming markedly greater in wheat compared to barley as weed density increases. Compared to wheat, barley limited yield and quality losses and suppressed intra-row weed growth more. Row spacing did not have a consistent effect on crop or weed parameters; in one of six experiments, the 25 cm row spacing reduced yields and increased intra-row weed biomass in wheat. Nitrogen rate did not affect crop or weed parameters. Results warrant the implementation of additional tactics to control intra-row weeds and limit crop losses.     

宁麦9号与扬麦158株高及其构成因素的遗传解析

宁麦9号与扬麦158是我国长江中下游麦区的主栽品种和骨干亲本,长江中下游麦区近3年来审定品种中80%都是其衍生后代,研究其性状的遗传具重要意义。以宁麦9号与扬麦158为亲本构建的包含282个家系的重组自交系群体为材料,利用Illumina 90k芯片对群体进行基因型分析,建立高密度遗传图谱。连续3个生长季对株高及节间长度、穗长等株高构成因素进行测定,结合遗传图谱对株高及相关性状进行QTL定位,获得14个控制株高及其构成因素的稳定表达位点。通过进一步位置比对,聚焦到6个染色体区段,初步明确了各节间对株高的遗传调控机制。同时,将6个染色体区段中同源性较低的连锁标记转化为适用于高通量筛选的KASP标记,利用101份区域试验材料进行标记效应验证,结果显示聚合Qph-2D与Qph-5A.1两个位点具有较高的选择效率,继续聚合Q2A后,中选材料显著减少,可能降低选择效率;对Q2A与Q5A两个一因多效位点的选择建议以降低株高的等位变异为主;Qd1-5D可作为穗下节间(D1)的选择标记对株高展开优化选择。期望以上结果能为长江中下游麦区的小麦株高遗传改良提供帮助。     

水稻白叶枯病菌对拌种灵抗药性分子机制研究

  通过紫外诱变获得了抗拌种灵水稻白叶枯病菌（Xanthomonas oryzae pv.oryzae）的突变体，这些突变体可以在含100 μg·ml-1拌种灵平板上生长，而敏感菌株在10 μg·ml-1浓度下则不能生长。敏感菌株琥珀酸脱氢酶活性受拌种灵强烈抑制，而抗药突变体的酶活性较低，且不受药剂抑制。应用6对引物，从水稻白叶枯病菌野生敏感菌株和室内诱导抗药性菌株中扩增到琥珀酸脱氢酶基因全序列。该基因全长3 616 bp，编码1 115个氨基酸，含有2个内含子。与柑橘溃疡病菌（Xanthomonas axonopodis pv. citri）琥珀酸脱氢酶核苷酸同源性93%，氨基酸同源性97%；与其它6种病原细菌琥珀酸脱氢酶基因编码的氨基酸序列同源性为43%～95%。敏感菌株和抗药菌株的琥珀酸脱氢酶序列分析表明，琥珀酸脱氢酶铁硫蛋白亚基中229位氨基酸由组氨酸（CAC）突变为酪氨酸（TAC）是导致X. oryzae对拌种灵产生抗药性的主要原因。     

Evaluation of the Pathotypes of Xanthomonas oryzae pv. oryzae and Preliminary Analysis of the Resistant Reactions of Main Japonica Rice in the Yunnan Plateau, China

The pathogenicity of 36 isolates of Xanthomonas oryzae pv. oryzae （Xoo）, which were collected from japonica rice varieties in the Yunnan Plateau, China, was evaluated. It was evaluated on 29 rice varieties including a set of seven varieties to identify pathogenicity, i.e., Haonuoyang, TN1, Kogyoku, Zhenzhu＇ai, IR26, Nanjing 33, and Kinmaze, which may be considered as a set of differential varieties for Xoo races from Yunnan japonica rice. The efficiency of the seven varieties was further confirmed. The results showed reversible and specific interactions between isolates and varieties. The isolates were classified into nine pathotypes from pathotyp Ⅰ to Ⅸ according to their pathogenic reactions on the seven rice varieties. The pathotype V was the epidemic, whereas pathogen Ⅶ was the most pathogenic. Most japonica varieties grown in the Yunnan Plateau were susceptible to Xoo. The rice lines IRBB21 （Xa-21）, Zhachanglong （Xa-22,, Xa- 24,）, and IR1545-339 （xa-5）, which were resistant to all the isolates tested, can be used as donors of resistant genes for bacterial blight in japonica rice breeding in the Yunnan Plateau.     

A-3中抗条锈新基因YrTp1和YrTp2的分子标记定位分析

【目的】半个多世纪的中国小麦育种史基本是育种家与条锈病的赛跑史。因此，筛选、鉴定、储备和利用新抗源是我国育种和资源研究中的一个长远战略性课题。【方法】利用小麦条锈菌条中31、32号生理小种，对来自小麦与十倍体长穗偃麦草［Thinopyrum ponticum (Host) Liu & Wang］的杂交后代材料A-3进行抗性遗传分析。用荧光SSR分子标记技术，鉴定所携带抗条锈病基因是否为新基因，并对其进行染色体定位研究。【结果】遗传分析表明，A-3对条中31号和32号的抗性由一显一隐2对基因控制。经过对196对微卫星引物的筛选，发现2B染色体短臂上的WMC477-167bp与显性基因紧密连锁，遗传距离为0.4 cM,将该显性基因定位于2BS上；7B染色体短臂上的WMC364-208bp与隐性基因连锁，遗传距离为5.8 cM。图位比较、系谱分析和抗谱分析表明，A-3所含抗条锈基因不同于已知抗条锈基因，暂定名为YrTp1和YrTp2。【结论】可利用A-3中与条锈病抗性紧密连锁的分子标记YrTp1和YrTp2将抗性基因转移到主栽品种中，在小麦育种和生产上发挥作用。