马铃薯种间体细胞杂种植株的细胞学观察和过氧化物同工酶谱分析

对马铃薯双单倍体品系81－15（2n＝2x－24）和南美二倍体栽培种Solanumphureja（2n＝2x＝24）体细胞融合获得的15个林系、81－15和二倍体野生种S．chacoense（2n＝2x＝24）体细胞融合获得的10个株系进行了细胞学鉴定和过氧化物同工酶谱分析。结果表明,杂种植株除一个株系为非整倍体外（染色体数为37）,其余株系均为四倍体（2n＝2x＝48）,是两个亲本的染色体数之和。与二倍体的双亲相比,杂种植株的叶下表皮气孔保卫细胞内叶绿体数目较多,但单位面积气孔数目较少,明显地表现出了四倍体的特征。杂种植株的过氧化物同工酶谱是其双亲酶谱谱带的总和。     

利用离体原生质体融合的方法获得马铃薯种间体细胞杂种

体细胞种间杂交在许多情况下获得了杂种植株和杂种试管植株。杂种植株基本上与有性双倍体相似,虽然有时具有染色体数目不同的特点。该项研究工作的任务在于研究马铃薯栽培种和野生种体细胞的杂交。在我们以往的著作中已经叙述了从得自分生组织的试管植株的叶肉分离原生质体、培养     

马铃薯种间体细胞杂种植株的细胞学观察和过氧化物同工酶谱分析

对马铃薯双单倍体品系８１－１５（２ｎ＝２ｘ＝２４）和南美二倍体栽培种Ｓｏｌａｎｕｍ ｐｈｕｒｅｊａ（２ｎ＝２ｘ＝２４）体细胞融合获得的１５个株系、８１－１５和二倍体野生种Ｓ．ｃｈａｃｏｅｎｓｅ（２ｎ＝２ｘ＝２４）体细胞融合获得的１０个株系进行了细胞学鉴定和过氧化物同工酶谱分析。结果表明,杂种植株除一个株系为非整倍体外９染色体数为３７）,其余株系均为四倍体（２ｎ＝２ｘ＝４８）,是两个亲本的染色体数之     

通过农杆菌介导把Harpin_(Ea)基因导入马铃薯的初步研究

应用农杆菌介导的叶盘法进行马铃薯遗传转化研究。通过D3、D4培养基筛选法 ,把HarpinEa蛋白基因导入马铃薯四倍体栽培种大西洋 (Atlantic) ,获得 10 7个转化株系。并经过卡那霉素的抗性鉴定和PCR分子检测 ,有 5 9株PCR检测呈阳性 ,占所有转化植株的 5 5 14%。结果表明HarpinEa基因已整合到马铃薯的基因组中。转化植株的抗病性鉴定正在进行中。     

马铃薯体细胞杂种主要形态性状和农艺性状鉴定

试验对来自马铃薯栽培种Solanum tuberosum与野生种S.chacoense原生质体融合产生的131个株系进行了形态学观察。结果表明,与亲本相比,大多数体细胞杂种的生长势较亲本弱,所有杂种的株高均小于融合亲本;匍匐茎长度均小于双亲平均数,其中小于栽培种亲本的体细胞杂种占42.1%;大多数体细胞杂种的叶片呈卵园形,叶形指数小于双亲。65%的体细胞杂种叶片有不同程度的褶皱现象。杂种群体的株高分离呈正态分布,匍匐茎长度的分离呈偏态分布。     

药用野生稻抗稻褐飞虱鉴定与利用技术研究

从198份药用野生稻资源中筛选出一批广谱高抗褐飞虱抗源（其中3份为免疫级）并对这些抗源进行了抗性遗传研究。研究结果表明其抗性是受一对显性基因控制的。通过离体幼胚培养获得绿苗,经过多代回交和自交,成功地将抗性基因转移到栽培稻中,获得了高世代（B4F5)株系。同时利用外源DNA花粉管导入法对药用野生稻广谱高抗褐飞虱基因导入到栽培稻进行了研究,获得了具有某些性状的后代。     

彩色马蹄莲品种对果胶软腐杆菌Pcc抗性的评价方法及鉴定

为探索彩色马蹄莲（Zantedeschia spp.）接种胡萝卜软腐果胶杆菌（Pectobacterium carotovora subsp. Carotovora, Pcc）的适宜方法,鉴定彩色马蹄莲组内品种对软腐病菌Pcc的抗性情况,本研究先以10个彩色马蹄莲品种为试验材料,进行以完整叶片、叶片圆片（叶盘）为接种材料的室内离体接种,以及以主脉、侧脉为接种部位的温室活体植株接种,接种后分别于12、24、36 h 3个时间点,以发病率、病情指数为指标,统计马蹄莲软腐病发病情况;在明确适宜的接种方法和发病时间后,选取包括第1步试验品种在内的34个彩色马蹄莲品种进行Pcc抗性鉴定。结果表明：离体接种的最佳材料为完整叶片,最佳发病观察时间是24 h;活体接种的最佳接种部位为叶片主脉,最佳发病观察时间是36 h;离体鉴定的中抗品种为5个,感病品种为11个,高感品种为18个;而活体鉴定的中抗、感病和高感品种分别为2、17、15个。离体方法鉴定的中抗品种多于活体鉴定,离体方法鉴定的抗性情况更接近田间大规模生产,其原因可能是离体方法鉴定时接种材料的均一性、环境条件的一致性好,更能反应品种本身的抗性情况。     

马铃薯体细胞杂种的青枯病抗性鉴定

本试验对马铃薯四倍体栽培种(Solanum tuberosum)的双单倍体系81-15与二倍体野生种S.cha-coense的原生质体融合株系进行了青枯病抗性鉴定,其目的是对利用体细胞融合技术获得抗青枯病种质的有效性及抗性鉴定技术进行评价,获得抗病育种材料。结果表明:在鉴定的18个体细胞融合株系中,抗性分离表现为从感病(S)到抗病(R),多数表现为中感(MS)到中抗(MR)水平。田间病圃鉴定、人工接种鉴定和分子标记鉴定的结果显示:田间病圃鉴定的病情指数(DI)和人工接种鉴定病级(DS)的相关性达到极显著水平(DI=-0.39+4 DS,r2=0.921),双侧翼SCAR标记SCA07446和SCA12980鉴定同时具有两个标记特异带的株系均为前两种鉴定表现为中感(MS)以上的材料。综合分析同时利用3种方法鉴定的结果:CHT-3、CHT-5、CHT-6、CHT-10、CHT-15等5个株系为抗病材料,证明体细胞融合是利用野生种质资源抗性的有效途径。     

利用细胞工程育种方法选育甜菜抗褐斑病品系初报

在接种于含有甜菜褐斑病菌毒素的选择培养基上 ,获得了甜菜抗褐斑病株系NRH -1的培养苗。于室内采用离体叶片法对这些培养苗进行了抗褐斑病性鉴定 ,结果表明 :与对照培养基(无毒)上获得的胚珠植株比 ,经含毒素培养基筛选的胚珠植株对褐斑病抗性较RH -1确有明显提高 ,70余株抗性植株 ,病级在2级以下的抗性植株占总株数21% ,为不同于原RH -1的抗病新株系。该方法有广阔的应用前景 ,为甜菜抗病育种提供一条新途径     

马铃薯种间体细胞杂种植株的形态学和农艺性状观察

对马铃薯双单倍体品系81－15和南美二倍体栽培种Solanumphureja原生质体融合获得的8块愈伤组织再生的15个株系、81－15和二倍体野生种S．chacoense原生质体融合获得的3块愈伤组织再生的10个株系的形态学特征和农艺性状进行了观察。结果表明，杂种植株的多数形态学特征和农艺性状，如分枝型、茎色、叶色、叶形、茎和茎叶绒毛、薯块皮色等呈现双亲的中间型，部分性状，如株型、花色等表现为偏亲性状。杂种植株与其双亲相比，生活力强，生长旺盛，叶片宽，块茎大。     

Resistance to soft rot (Erwinia carotovora subsp.atroseptica) in tetraploid potato families obtained from 4x-2x crosses

Diploid potato clones, interspecific hybrids ofSolanum species, having in their originS. tuberosum,S. chacoense, S. yungasense, S. phureja, S. gourlayi, andS. demissum, with resistance to soft rot, were crossed to tetraploid potato clones in 4x-2x crosses. The 24 tetraploid families obtained in a North Carolina II design were examined for tuber resistance to soft rot in a laboratory test and for basic agronomic traits in field trials conducted for two consecutive years. In addition, one family originating from a 4x-2x cross of two susceptible parents was tested. Statistical analysis of the data revealed significant general (GCA) and specific combining ability (SCA) effects, year, GCA (female) × year, GCA (male) × year, and SCA × year upon the inheritance of resistance to soft rot. About 35% of the progeny was selected as resistant to tuber soft rot, and of these 11% showed high resistance combined with good tuber yield, tuber weight, and tuber appearance. The relationships between resistance to soft rot and chosen agronomic traits were not noted or were weakly significant and sporadic. The resistance to tuber soft rot found in diploid potato hybrids can be transferred to the cultivated tetraploid pool through 4x-2x crosses, and a high frequency of offspring posses resistance.     

Introgression and stabilization oferwinia tuber soft rot resistance into potato after somatic hybridization ofsolanum tuberosum ands. brevidens

Resistance to potato tuber soft rot caused byErwinia carotovora was transferred fromSolanum brevidens to the cultivated potato over the course of four backcross generations originating from a somatic hybrid. Soft rot reactions were determined via a tuber plug inoculation method developed during the course of these experiments. Soft rot resistance was highest in the somatic hybrid (only ca. 20% of tubers and plugs showed evidence of severe rotting) and lowest among progeny of control potato x potato crosses (ca. 80% of tuber plugs showed severe rot). Backcross generations involving somatic hybrids were intermediate in their reaction, and resistance stabilized to about 60% of tuber plugs showing severe rot in the BC₂ through the BC₄. Reciprocal crosses showed no difference in the inheritance of soft rot resistance, indicating that neitherS. brevidens norS. tuberosum donor cytoplasm had a significant effect on the expression of resistance. Crosses between BC₃ siblings where noS. brevidens genetic markers were detected but resistance was segregating demonstrated a dosage effect for soft rot resistance. We conclude that introgression of soft rot resistance has occurred and that at least one locus responsible for resistance inS. brevidens now resides in theS. tuberosum genome.     

The inheritance of resistance to soft rot (Erwinia carotovora subsp.atroseptica) in diploid potato families

Soft rot of potato tubers, caused byErwinia (Pectobacterium), is a serious disease affecting potato crops during storage. Studies on the inheritance of resistance to tuber soft rot were undertaken on diploid potato hybrids. A total of 480 clones, derived from 12 families, were examined for resistance to soft rot in laboratory tests over 3 years and for basic agronomic traits in field experiments over 2 years. General (GCA) and specific (SCA) combining abilities were significant in the inheritance of resistance to soft rot; however, GCA for female parents were significant in 1 or 2 years during 3 years of evaluation and SCA was significant for eight, two, and six families out of 10 tested in 1994, 1995, and 1996, respectively. There were also significant differences between years. Moreover, marked interactions for GCA_females × year and SCA × year in the variation of soft rot resistance were found. Broad-sense and narrowsense heritability of resistance, measured as diameter of rotten tissue, was estimated as 0.92 and 0.89, respectively. The maternal effect, evaluated in two sets of reciprocal crosses, was not significant for the genetic determination of resistance to soft rot. No significant relationships were found between resistance to soft rot and the main agronomic traits in the tested diploid families. These results suggest that diploid potato clones with resistance to soft rot can be selected after being evaluated over a few seasons. It is also possible to select clones combining good resistance to soft rot with high yield, superior tuber characteristics, and acceptable starch content. These resistant diploids can be used as male parents in 4x-2x crosses in breeding tetraploid potato resistant to soft rot.     

Soft rot and blackleg reactions in diploid potato hybrids inoculated withErwinia spp.

From 1993 to 1996 three groups of potato genotypes were evaluated for resistance toErwinia spp.: (1) 31 interspecific diploid hybrids (28 resistant and three susceptible), (2) five hexaploid or pentaploid somatic hybrids ofSolanum tuberosum (tbr) ×S. brevidens (brd), and (3) eight cultivars. Two evaluation methods were applied: tuber point inoculation with eitherErwinia carotovora ssp.atroseptica (Eca) orE. chrysanthemi (Ech) to test tuber soft rot resistance and stem base inoculation with Eca to test blackleg resistance. Some resistant diploid hybrids and somatic hybrids oftbr × brd were superior to cultivars for both tuber and stem resistance. Tuber and stem resistance toErwinia spp. in the most resistant diploid hybrids were equal to the highly resistant somatic hybrids oftbr xbrd. Tuber resistance to Eca was highly correlated to tuber resistance to Ech (r=0.815***). In two years of evaluation for stem resistance, three diploid hybrids and a derivative of one of the somatic hybrids (USA M 264) failed to develop symptoms of blackleg following inoculation with Eca, Analyses of variance for tuber and stem resistance indicated significant effects of genotype, year and genotype × year interaction. A positive relationship between tuber and stem resistance toErwinia spp. has been found, however the genetic control of resistance in tuber and stem is partially independent. In the case of Eca the correlation coefficient was r = 0.725***. Therefore it should be possible to obtain resistant genotypes to both blackleg and tuber soft rot. Several resistant diploid hybrids were selected from among those tested, which also have several other characters desirable for potato breeders.     

Resistance to blackleg and tuber soft rot in sexual and somatic interspecific hybrids with different genetic background

Diploid interspecific hybrids from crosses betweenSolanum tuberosum haploids andS. canasense, S. multidissectum, andS. tarijense, along with tetraploid and hexaploid somatic hybrids S.tuberosum (+)S. commersonii, were screened for resistance to blackleg and tuber soft rot byErwinia carotovora subsp.atroseptica andE. carotovora subsp.carotovora. Among the diploid hybrids, those involving clone mlt la of S.multidissectum and tar 2b ofS. tarijense yielded the highest number of genotypes resistant to both blackleg and tuber soft rot. By contrast, all the hybrids involving clone tar 1lb ofS. tarijense were susceptible to both bacterial diseases. As far as resistance of somatic hybrids is concerned, the most interesting genotype was the tetraploid one which showed resistance to both blackleg and tuber soft rot Significant correlations were found betweenE. carotovora subsp.atroseptica andE. carotovora subsp.carotovora both in resistance to blackleg and in resistance to tuber soft rot, suggesting that resistance to the two subspecies is controlled by the same gene/s or that the relevant genes are linked. On the other hand, correlations between blackleg and tuber soft rot were never significant, indicating that different mechanisms may control resistance in tuber and stems. Alternatively, the resistance could be controlled by the same gene/s under a different spatial/temporal expression pattern. Twelve diploid sexual hybrids and one tetraploid somatic hybrid were selected for resistance to blackleg and/or tuber soft rot and for tuber characteristics.     

Erwinia resistance in backcross progenies ofSolanum tuberosum x S. tarijense andS. tuberosum (+) S. commersonii hybrids

Summary The resistance to tuber soft rot caused byErwinia carotovora ofSolanum tuberosum x S. tarijense andS. tuberosum (+) S. commersonii hybrids and their backcrosses is reported. A number of resistant diploid sexual hybrids and tetraploid/hexaploid somatic hybrids were selected. Backcross progenies were obtained through 2x×4x crosses involving a resistant diploid hybrid and tetraploidS. tuberosum, and through 4x×4x crosses between a resistant somatic hybrid andS. tuberosum. The hybrids showed high variability interms of resistance to tuber soft rot. The resistance of progeny from 2x×4x backcrosses was similar to that of the parental sexual hybrid. By contrast, the resistance of genotypes deriving from 4x×4x backcrosses was reduced compared with the resistant somatic hybrid. In general, tuber characteristics of the backcross hybrids improved considerably as compared with their parents, and tuber yield per plant was good.     

Breeding Potential of Potato Somatic Hybrids: Evaluations for Adaptability,Tuber Traits,Late Blight Resistance,Keeping Quality and Backcross (BC<Subscript>1</Subscript>) Progenies

Ten tetraploid interspecific potato somatic hybrids developed earlier through protoplast fusion between the dihaploid Solanum tuberosum L. ‘C-13’ and the diploid wild species Solanum pinnatisectum Dun. were used in this study. Somatic hybrids and standard control cultivars were evaluated for adaptability, tuber traits, late blight resistance and keeping quality attributes (dormancy, sprouting, weight loss and appearance after 75 days of storage) during two successive winter crop seasons (short-days) in the sub-tropical plains of India, where nearly 90% of India’s potatoes are grown. Somatic hybrids showed medium to good plant vigour and late to very late plant maturity. Though none of the somatic hybrids produced equal or higher tuber yield than the control cultivars, except clone P7, all other somatic hybrids produced significantly higher marketable and total tuber yield after a 90-day growth than the parent C-13. Most of the somatic hybrids possessed higher tuber dry matter concentration, resistance to late blight and better keeping quality attributes than the control cultivars. Notably, the tubers of somatic hybrids showed a tendency for colour change from white to purple on exposure to sunlight. Breeding potential of somatic hybrids was further assessed by crossing them with commonly grown local potato cultivars for the development of backcross (BC₁) progenies. In the hybridization programme, berries were formed while somatic hybrids were used either as male or female parents with common cultivars. Our results revealed that the use of bulk pollen of somatic hybrids not only resulted in higher pollination success but also helped to achieve higher numbers of true potato seeds (TPS) per berry. Despite the formation of berries, TPS was absent in 22 crosses using somatic hybrids as a female or male parent which may be due to ovule or embryo abortions. Segregating BC₁ generations were evaluated under the same sub-tropical conditions for further exploitation in potato breeding. Based on our study, the somatic hybrids P4, P8 and P10 can be utilized for the introgression of important characters such as high tuber dry matter concontration, resistance to late blight and excellent keeping quality attributes into the cultivated potato via conventional breeding methods for cultivar development in the sub-tropical plains of India.     

Sexual progeny of somatic hybrids between potato andSolanum brevidens: Potential for use in breeding programs

Hexaploid somatic hybrids have been obtained by fusion of protoplasts fromSolanum brevidens (PI 218228, 2x = 2n = 24) andS. tuberosum (PI 203900 or cv. Russet Burbank; 2x = 4n = 48). In the work reported here, pentaploid progeny derived from sexual crosses between the somatic hybrids and the potato cultivar, Katahdin were assessed for transfer of disease resistances and improvement of agronomic traits. Segregation was noted in disease resistances to race 0 ofPhytophthora infestans and to tuber soft rot caused byErwinia spp., indicating that sexual transfer of traits captured by protoplast fusion had occurred. The pentaploid progeny showed substantial variation in and often improvement of some agronomic traits as compared to the somatic hybrids. The results indicate that interspecific somatic hybridization can produce material with potential for use in a potato breeding program     

Inheritance of a pectate lyase mediated soft rot resistance in crosses between transgenic potato lines of cv. désirée andS. tuberosum cultivars

Summary Expression of the pectate lyase (PL) isoenzyme 3 in transgenic potato lines of cv. Désirée mediates an enhanced resistance of tuber tissue toErwinia carotovora (Ec) soft rot due to a pre-activation of plant defence mechanisms. Therefore, theSolanum tuberosum cvs Agave and Adretta with a moderate level of soft rot resistance were crossed with such PL-expressing potato lines. The resulting progenies were assessed with respect to plant/tuber characteristics over a period of four years and then tested for PL3-expression as well as for soft rot resistance. 71% of the selected progeny lines exhibited a stable production of the PL3 enzyme. Statistical analysis revealed differences between the transgenic and the non-transgenic progeny concerning the soft rot resistance of tuber tissue. Compared with the PL-inactive progeny, extension of Ec-rotting on the wound surface of PL-transgenic potatoes was diminished on average by 51.8%. Similarly, the degree of cell lysis caused by bacterial maceration was significantly (P<0.05) reduced in tuber tissue of PL-expressing progeny lines. The latter also revealed an enhanced PPO and PAL activity in their tuber tissue indicating an active plant defence. It is concluded therefore that the PL-mediated soft rot resistance introduced into potatoes by means of molecular techniques is heritable.     

Frost resistant potato hybrids viaSolanum acaule,Bitt. Diploid-Tetraploid crosses

Solanum juzepczukii Buk. andSolanum curtilobum Juz. et Buk. are two important primitive cultivated species of potato which are planted in the highlands of Peru and Bolivia. Little natural variation exists within these species. Crosses were made to artificially recreate these species using tetraploid cultivated potatoes as female parents and triploids derived from acaule-diploid crosses as males. The new hybrids were tetraploid (2n = 48) with a high degree of heterosis and variability, and several potential cultivated clones possessing valuable characters such as yield, earliness, frost tolerance and good tuber type were selected.