玉米单倍体DH系特点及应用研究进展

利用单倍体诱导系选育技术能显著加快育种进程,高效单倍体诱导材料的培育及单倍体DH系应用是玉米单倍体育种的重要基础.本文就玉米单倍体DH系的特性及应用加以阐述,并对今后玉米单倍体DH系研究进行了展望.     

玉米单倍体加倍(DH)系籽粒性状表现的研究

玉米单倍体育种是快速选育纯合自交系的方法.研究利用单倍体诱导系,对来自BSSS杂种优势群的两个自交系BX08和BX19的杂交F1进行诱导获得单倍体,又经加倍获得33个加倍纯合系(DH),分析单倍体加倍纯合系与两个亲本及F1杂交种的果穗及籽粒性状表现.结果表明,单倍体加倍系的籽粒容积和质量比亲本籽粒都有所提高,子代的农艺性状获得显著改善,说明发生单倍体不是基因随机分离的结果,单倍体育种存在选择压力,有利于优良基因的累积,淘汰不良基因.     

玉米单倍体诱导系诱导性状的杂种优势分析

以8个玉米单倍体诱导系和按照NCⅡ方法进行杂交组配的28个诱导系F1为材料,分析株高、雄稳小花数的杂种优势,并利用6个骨干自交系京724、京92、京X 005、昌7-2、郑58和农系531为测验种进行单倍体诱导率测验,分析诱导率的杂种优势.结果表明,25个诱导系F1的株高表现出高亲优势,最高超高亲36.7 cm;28个诱导系F1的雄穗小花数都表现出高亲优势,最高超高亲265.4个;15个诱导系F1诱导率表现出高亲优势,最高超高亲2.30％;综合诱导率、株高、雄穗小花数3个诱导性状分析,表现优良的组合有JKY 042/JKY 044、JKY043/JKY 044、JKY 044/JKY 045、JKY 042/JKY 046、JKY 042/JKY 043.因此通过正确选用不同来源诱导系进行杂交组配,可综合利用诱导率、株高、雄穗小花数的杂种优势,用于单倍体DH工程化育种进程中的规模化隔离诱导.     

同一来源玉米DH系的性状分离分析

利用单倍体诱导系对遗传背景相近的玉米自交系MC1和MC2所组配的F1进行杂交诱导获得单倍体,经过自然加倍得181份纯系(DH系),分析DH系农艺性状的分离情况。结果表明:181个DH系株高、穗位高、穗长、秃尖长、穗行数、百粒重和单穗粒重等数量性状的次数分布符合正态分布;所得DH系各数量性状的变异类型丰富,且在产量性状上存在优于两亲本的分离;两亲本的21对SSR核心引物分子标记带谱在DH系中的分布都近似于1∶1比率,未出现偏分离。因此,通过遗传诱导的方法所得DH系类型多样,可以满足育种上的需要。单倍体育种是一种选育优良玉米纯系的有效方法。     

不同诱导系对不同基因型玉米材料的诱导率评价

玉米双单倍体（DH，Doubled haploid）技术是现代玉米育种的关键核心技术，能显著加快育种进程。为解决目前玉米单倍体育种普遍存在的诱导率低、杂株率高等问题，本研究利用 4 个单倍体诱导系和 12 个基础群体材料进行杂交，探讨不同诱导系及不同基因型材料对单倍体诱导率的影响。结果表明：（1）用 4 个诱导系诱导同一基础材料时 CS3 诱导系的平均诱导率最高，为 5.66%，稳定性也较好；不同基础材料与同一诱导系杂交时 D3 和 D4 的平均诱导率最高，分别达 7.44% 和 7.45%。 （2）用 4 个诱导系诱导同一基础材料时 CS3 诱导系的杂株率最小，为 16.95% ；不同基础材料与同一诱导系杂交时 HB4 的平均杂株率最小，为 10.93%。（3）诱导系 CS3 对 HB4 诱导时诱导率最高（11.9%），杂株率最低（0.81%）。研究结果可充实玉米单倍体育种理论研究，为玉米单倍体育种生产奠定理论基础。     

玉米DH系的表现及其同工酶电泳分析

以孤雌生殖诱导系高诱1号诱导的单倍体植株为材料,由单倍体加倍后获得的DH系性状上表现高度一致,其中一些DH系的长势和果穗性状达到了育种上的要求.DH系的过氧化物酶和酯酶同工酶电泳表明,部分DH系具有母本谱带,部分具有父本谱带,由此说明,孤雌生殖诱导系诱导单倍体具有一定的随机性.     

我国玉米地方种质的单倍体诱导和加倍特性研究

双单倍体(doubled haploid,DH)育种技术具有加速育种进程的突出优势,已成为玉米育种关键性核心技术并在国外广泛应用.本试验选用47份玉米地方种质,进行单倍体诱导和单倍体加倍特性研究.结果表明,47份玉米地方种质材料之间杂交诱导的拟单倍体率有显著差异,介于1.64％～14.50％之间,平均为5.95％.种植12份玉米地方种质的拟单倍体籽粒进行田间鉴定,标记鉴定准确率介于40.2％～82.3％之间,校正单倍体诱导率介于2.33％～6.45％之间,表明玉米遗传背景影响到籽粒标记的表达,单倍体诱导率有明显差异.将15份玉米地方种质拟单倍体于冬季在海南田间种植,加倍授粉株率介于2.7％～27.2％之间,加倍结实株率介于0.7％～8.9％.说明我国玉米地方种质的遗传多样性丰富,单倍体诱导率和加倍率具有明显差异,利用DH育种技术可以拓宽和加强我国玉米地方种质在玉米育种中的利用.     

孤雌生殖诱导玉米单倍体育种技术研究

玉米单倍体育种技术可以有效地缩短育种进程,快速获得大量纯合系。孤雌生殖诱导是获得玉米单倍体的主要方法之一,该项技术在国外各大育种公司已被广泛使用,但在国内仍处于研究探索阶段。本文从诱导系的选择与繁殖、基础材料的组配、杂交诱导、单倍体籽粒的挑选、单倍体植株的种植、加倍以及双单系的繁殖等方面综述了孤雌生殖诱导系（简称诱导系）产生玉米单倍体育种技术的基本流程,同时展望了单倍体育种的应用前景。     

小麦×玉米远缘杂交技术以及在单倍体育种上的应用评价

为了提高小麦作物育种选择的精确性和效率,应用小麦亲本杂交后代F1(或F2)代与玉米进行远缘杂交,经激素诱导和组织培养形成单倍体胚及植株,加倍单倍体(DH)育种结合分子标记和性状表达,达到了单代纯化培育小麦新品系的生产能力;本文阐述了小麦×玉米远缘杂交发展进程,分析了小麦×玉米远缘杂交较孤雄生殖、球茎大麦技术等具备的优势...     

玉米单倍体诱导系Stock6及其在玉米育种中的应用

玉米单倍体育种可缩短育种周期、提高育种效率,利用玉米单倍体诱导系诱导玉米单倍体简单易行,单倍体筛选方便。简述了玉米单倍体诱导系Stork6的发现、改良及在玉米育种中的应用。     

DH群体内株系对籼、粳亚种的亲和性

以窄叶青8号（ZYQ8）/京系17(JX17)的13个花培DH株系和培矮64／02428的11个花培DH株系为材料,分析了它们的籼粳分化及其对籼、粳亚种杂交的亲和性。结果表明,无论亲本是否具有广亲和性亲缘,两个DH群体均出现广亲和性良好的株系,且多数株系属籼粳中间类型。这说明虽然广亲和性是遗传的,但是籼粳基因的重组确实可以产生广     

Segregation distortion in doubled haploid lines of barley (Hordeum vulgare L.) detected by simple sequence repeat (SSR) markers

A total of 147 simple sequence repeat (SSR) markers (including86 barley and 61 wheat microsatellite markers) were tested for their segregation in a doubled haploid (DH) and an F₂ population of barley. The DH population consisted of 71 doubled haploid lines, developed from F₁ plants of a cross between Tadmor and WI2291using isolated microspore culture technique. A genetic linkage map consisting of 43 microsatellite markers was constructed using the DH population. Particularly on chromosome 4H microsatellite markers showed distorted segregation ratios. Segregation of DH lines based on molecular markers were compared with segregation of 92 F₂ lines from the same cross. The proportion of loci deviating from the expected monogenic segregation ratios in the DH population was significantly higher (19/43loci, 44%) than in the F₂ population (7/43 loci, 16%). The deviation was biased towards the WI2291 parent alleles. In line with this observation, WI2291 was found to perform better than Tadmor in regenerating green plantlets with the isolated microspore-culture technique. This revised version was published online in July 2006 with corrections to the Cover Date.     

水稻叶鞘相关性状的遗传分析

本研究以典型釉粳交(春江06/台中本地1号)的F1代花药加倍的DH群体为材料,系统考察了DH群体及其双亲在抽穗期的叶鞘长、叶鞘重、叶鞘厚等性状上的分布情况.结果表明,叶鞘长度的遗传力较高,受环境的影响相对较小,叶鞘长与叶鞘厚及叶鞘干重间大多呈极显著的正相关.QTL分析共检测到31个与叶鞘有关的QTL,分布于水稻第1、第...     

Haploid plantlet screening in the development of blackleg resistant DH lines of Brassica napus

This study was conducted to expedite disease-resistance breeding in canola by screening haploid plants against blackleg disease. Microspore-derived haploid plantlets from 14 unrelated crosses were inoculated with Leptosphaeria maculans pycnidiospores and were rated as resistant, intermediate or susceptible. Blackleg-inoculated and control haploid plants were then colchicine-treated to produce doubled haploid (DH) lines. The DH lines thus produced were again screened against blackleg using the cotyledon bioassay. In general, the proportion of the resistant DH lines derived from selected resistant haploid plants was much higher than the proportion of resistant DH lines from control haploid populations. There was no detrimental effect on the survival of haploid plants after the combined treatment of blackleg inoculation and colchicine. The importance of the micro-environment on the final outcome of plant-pathogen interaction in case of non-obligate pathogens is also discussed.     

Discrimination of haploid and diploid maize kernels via multispectral imaging

The use of doubled haploids (DHs) in maize has become ubiquitous in maize breeding programmes as it allows breeders to go from cross to evaluation in as little as 2 years. Two important aspects of the in vivo DH system used in maize are as follows: (i) the identification of haploid progeny and (ii) doubling of the haploid genome to produce fertile inbred lines. This study is focused on the first step. Currently, identification of maize haploid progeny is performed manually using the R1‐nj seed colour marker. This is a labour‐intensive and time‐consuming process; a method for automated sorting of haploids would increase the efficiency of DH line development. In this study, six inbred lines were crossed with the maternal haploid inducer ‘RWS/RWK‐76’ and a sample of seed was sorted manually for each line. Using the VideometerLab 3 system, spectral imaging techniques were applied to discriminate between haploids and hybrids. Using DNA markers to confirm the haploid/diploid state of the tested seed, for the majority of genotypes haploid identification was possible with over 50% accuracy.     

Classification approaches for sorting maize (Zea mays subsp. mays) haploids using single-kernel near-infrared spectroscopy

Doubled haploids (DHs) are an important breeding tool for creating maize inbred lines. One bottleneck in the DH process is the manual separation of haploids from among the much larger pool of hybrid siblings in a haploid induction cross. Here, we demonstrate the ability of single-kernel near-infrared reflectance spectroscopy (skNIR) to identify haploid kernels. The skNIR is a high-throughput device that acquires an NIR spectrum to predict individual kernel traits. We collected skNIR data from haploid and hybrid kernels in 15 haploid induction crosses and found significant differences in multiple traits such as percent oil, seed weight, or volume, within each cross. The two kernel classes were separated by their NIR profile using Partial Least Squares Linear Discriminant Analysis (PLS-LDA). A general classification model, in which all induction crosses were used in the discrimination model, and a specific model, in which only kernels within a specific induction cross, were compared. Specific models outperformed the general model and were able to enrich a haploid selection pool to above 50% haploids. Applications for the instrument are discussed.     

pinⅡ基因在转基因大白菜DH纯系和试配F1代的遗传及表达

马铃薯蛋白酶抑制剂基因-Ⅱ(pinⅡ)是一类控制丝氨酸蛋白酶抑制剂PI-Ⅱ合成的抗虫基因.为了比较pinⅡ基因在转基因大白菜纯系和杂种中的表达特性,为抗虫新品种选育提供理论指导,本实验以转基因大白菜双单倍体纯系(DH系)的5个自交系和与常规育种亲本材料试配的3个F1杂交组合为试材,利用标记性状(除草剂抗性)对试材进行转...     

玉米高频率孤雌生殖单倍体诱导系的选育与鉴定

本试验从单倍体诱导系Stock6与高油玉米群体BHO的杂交后代中经过不断的测交和选择, 育成了我国第一个孤雌生殖单倍体诱导系农大高诱1号。 在16个测交组合中高诱一号对411自交系的单倍体诱导率最高(9.2%); 而对H4自交系的单倍体诱导率最低(1.92%); 其平均单倍体诱导率为5.34%。 以黄绿苗自交系Syn695yg为测验种, 高诱1号的     

籼粳交加倍单倍体后代性状遗传的研究

对一个典型的籼/粳组合、窄叶青8号/京系17的F1花药进行培养,所获二倍体花粉植株除约20%自然结实率为0外,其余植株的结实率在1%至正常结实之间,65%株系在75%以上,并稳定地传至自交后代。对130多个加倍单倍体(DH)多种性状的遗传进行了研究与分析,结果表明,花药培养可以克服籼/粳后代包括育性在内的各种性状长期分离不易     

Variation in Doubled Haploid Plants of Wheat Obtained through Wheat (Triticum aestivum) × Maize (Zea mays) Crosses

Variation was investigated in 110 doubled haploid (DH) lines of wheat derived from wheat × maize crosses. Field observation revealed visible variations in 15 DH2 lines such as extreme dwarfism, low seed fertility, alteration of spike type and stripes. Six agronomic traits, i.e., heading date, spike number/ plant, culm length, spike length, seed fertility and grain weight were statistically analyzed in the DH2 and DH3 generations. Out of the 88 DH2 lines/DH3 groups, 26 %/64 % showed significant differences from the parental variety in the means of one or more traits. Ranges of the DH3 lines were larger than those of the DH2 lines, except for spike number/plant. Furthermore, analyses of variance within and between DH lines showed the presence of heterogeneity/heterozygosity in the DH2 lines/plants. These results indicated the occurrence of gametoclonal variation in the DH lines. It is considered that most of the variations detected were due to the colchicine treatment rather than to the 2,4-D treatment or in vitro culture.