青Qian愈伤组织在继代培养中的分化能力及染色体稳定性研究

青Ｑｉａｎ胚性愈伤组织在改良５９附加２,４－Ｄ及Ｋｔ及１ｐｐｍ的培养基上继代３年（每月继代１次）,仍具有旺盛的增殖能力,在胚性愈伤组织转入１／２改良５９并附加ＡＢＡ１ ｐｐｍ的分化基上,约３个月左右可分化出大量体细胞胚,体细胞胚分化率达０％以上,经继代３年的胚性愈伤组织细胞的染色体倍性十分稳定,其染色体数及核型为２ｎ＝２４＝１６ｍ（６ｓｃ）＋８ｓｍ＋２Ｂ。这一结果与由实生苗根尖压片所得结果基本一致     

白Qian胚性愈伤组织长期继代培养中的分化能力及染色体稳定性 …

白Ｑｉａｎ（Ｐｉｃｅａ ｍｅｙｅｒｉ Ｒｅｈｄ．ｅｔ．Ｗｉｌｓ．）的胚性愈伤组织ＭＳ＋２,４－Ｄ１ｍｇ／Ｌ＋ＫＴ１ｍｇ／Ｌ的培养基上继代３年,增殖能力和分化潜力仍保持在原来的水平,没有明显降低的趋势。但随着继代时间的增长,胚性愈伤组织内有细胞的染色体数目发生了无规律的变化,而再生植株根尖细胞染色体数目比较稳定（２ｎ＝２８）。     

青扦胚性细胞悬浮培养中影响体细胞胚发生因素的研究

试验以青扦（Ｐｉｃｅａｗｉｌｓｏｎｉ）的胚性愈伤组织为材料,以改良５９基本成分附加２４－Ｄ１ｍｇ／Ｌ及ＫＴ１ｍｇ／Ｌ为培养介质,比较了液体悬浮与半固体二种培养方式对胚性愈伤组织增殖和体细胞发生的影响,研究了液体悬浮培养过程中影响体细胞胚发生的因素。结果表明：液体悬浮培养好于半固体培养,它的胚性愈伤组织的生长率为２６８％,是半固体培养的１２４倍;体细胞胚的分化率为９３％,是半固体培养的２２倍;悬浮培养较佳的培养条件为：初始细胞密度为２％（鲜重）,蔗糖浓度为２０ｇ／Ｌ,摇床转速为１００ｒ／ｍｉｎ,ｐＨ为５８。经过两个月悬浮培养,将培养物转至１／２改良５９附加ＡＢＡ１ｍｇ／Ｌ的分化培养基上,３个月后每ｇ培养物上可获得２８５个正常的子叶期体细胞胚。     

白皮松和油松雌配子体愈伤组织的诱导和分化

以白皮松（Ｐｉｎｕｓ ｂｕｎｇｅａｎａ Ｚｕｃｃ．）和油松（Ｐ．ｔａｂｕｌａｅｆｏｒｍ ｉｓ Ｃａｒｒ．）的未成熟胚乳,即雌配子体为外植体进行培养,将雌配子体分别接种在添加不同激素种类和不同浓度配比的改良ＭＳ培养基上诱导愈伤组织。经过２０多天的培养,在含有１—６ ｍ ｇ／Ｌ 萘氧乙酸（ＮＯＡ）和０．５ｍ ｇ／Ｌ６－ＢＡＰ及３％ 蔗糖浓度的培养基上诱导产生了愈伤组织,愈伤组织的诱导频率最高为２５％ 。经细胞学观察证明：愈伤组织细胞确为单倍性的,染色体数目为ｎ＝ １２,正常的体细胞染色体数目为２ｎ＝ ２４,并在含有ＡＢＡ 的原诱导愈伤组织培养基上分化出绿色小芽     

山东10种植物的核型分析

对山东１０ 种植物进行了核型分析。茴茴蒜（ Ｒａｎｕｎｃｕｌｕｓｃｈｉｎｅｎｓｉｓ Ｂｇｅ－） 染色体数目２ｎ ＝１６ , 核型公式Ｋ（２ｎ） ＝ ２ｘ ＝ １６ ＝ ２ Ｍ ＋ ２ｍ ＋ ２ｓｍ ＋ １０ｓｔ, “３Ａ”类型; 五脉地椒（ Ｔｈｙｍｕｓｑｕｉｎｑｕｅｃｏｓｔａｔｕｓ Ｃｅｌａｋ－） 染色体数目２ｎ＝ ２６ , 核型公式Ｋ （２ｎ） ＝ ２ｘ＝ ２６ ＝ ８ Ｍ ＋ １８ｍ , “１Ａ”类型; 蛇床（ Ｃｎｉｄｉｕｍ ｍｏｎｎｉｅｒｉ（Ｌ－） Ｃｕｓｓ－） 染色体数目２ｎ＝ ２０ , 核型公式Ｋ （２ｎ） ＝ ２ｘ＝ ２０ ＝ ２Ｍ＋ １６ｍ ＋ ２ｓｍ , “２Ｂ”类型; 波斯菊（ Ｃｏｓｍｏｓ ｂｉｐｉｎｎａｔｕｓ Ｃａｖ－） 染色体数目２ｎ ＝ ２４ , 核型公式Ｋ（２ｎ） ＝ ２ｘ ＝ ２４ ＝ １６ｍ ＋ ２ｍ （ｓａｔ） ＋ ６ｓｍ , “２Ａ”类型; 白车轴草（ Ｔｒｉｆｏｌｉｕｍ ｒｅｐｅｎｓ Ｌ－） 染色体数目２ｎ＝ ３２ , 核型公式Ｋ （２ｎ） ＝ ４ｘ ＝ ３２ ＝ ３２ｍ , “１Ａ”类型; 铁苋菜（ Ａｃａｌｙｐｈａ ａｕｓｔｒａｌｉｓ Ｌ－）染色体数目２ｎ ＝ ３２ , 核型公式Ｋ （２ｎ） ＝ ２ｘ＝ ３２ ＝ ３２ｍ , “１Ｂ”类型; 地构叶（ Ｓｐｅｒａｎｓｋｉａ ｔ?     

一品红离体培养诱导体细胞胚的研究（简报）

一品红的嫩茎离体培养在附加不同浓度的２,４－Ｄ和６－ＢＡ的ＭＳ培养基上,２．４Ｄ浓度为１．０～２．０ ｍｇ· Ｌ~（－１）的愈伤组织生长量最大。这些愈伤组织转移到ＭＳ＋６－ＢＡ１．０ ｍｇ·Ｌ~（－１）＋ＮＡＡ０．１ ｍｇ·Ｌ~（－１）的分化培养基上后,产生了胚性愈伤组织,观察到大量不同时期体细胞胚。带有体细胞胚的愈伤组织进一步转移到无激素 ＭＳ培养基上,便发育成小植株。     

刺槐宽叶和四倍体无性系的组织培养

１植物名称刺槐（Ｒｏｂｉｎｉａｐｓｅｕｄｏａｃａｃｉａ）优良无性系：Ｔｅｔｒａｐｌｏｉｄｌｏｃｕｓｔ、Ｇｌｇａｓｔｙｐｅｌｏｃｕｓｔ。２材料类别带腋芽的茎段。３培养条件（１）启动培养基：ＭＳ＋６－ＢＡ０．２５ｍｇ·Ｌ－１（单位下同）＋ＮＡＡ０．０５。（２）分化培养基和继代培养基：ＭＳ＋６－ＢＡ０．５＋ＮＡＡ０．１＋ＡｇＮＯ３１０,ＭＳ＋６ＢＡ０．５＋ＮＡＡ０．１。上述培养基均添加３％蔗糖、０．６％琼脂。（３）生根培养基：１／２ＭＳ＋ＩＢＡ０．２＋ＮＡＡ０．２,添加２％蔗糖０．６％琼脂。培养基ｐＨ…     

国产沙参属五个种的核型研究

首次报道了５种国产沙参属（Ａｄｅｎｏｐｈｏｒａ）植物的染色体数目和核型。北方沙参Ａ．ｂｏｒｅａｌｉｓＨｏｎｇｅｔＹ．Ｚ．Ｚｈａｏ的核型公式为２ｎ＝３４＝２８ｍ＋４ｓｍ（２ｓａｔ）＋２ｓｔ（２ｓａｔ）;雾灵沙参Ａ．ｗｕｌｉｎｇｓｈａｎｉｃａＨｏｎｇ的核型公式为２ｎ＝３４＝２６ｍ（４ｓａｔ）＋６ｓｍ＋２ｓｔ或２ｎ＝３４＋１Ｂ＝２６ｍ（４ｓａｔ）＋６ｓｍ＋２ｓｔ＋１Ｂ;秦岭沙参Ａ．ｐｅｔｉｏｌａｔａＰａｘｅｔＨｏｆｍ．的核型公式为２ｎ＝３４＝２６ｍ（２ｓａｔ）＋６ｓｍ＋２ｓｔ或２ｎ＝３４＋１Ｂ＝２６ｍ（２ｓａｔ）＋６ｓｍ＋２ｓｔ＋１Ｂ;裂叶沙参Ａ．ｌｏｂｏｐｈｙｌａＨｏｎｇ的核型公式为２ｎ＝３４＝２６ｍ＋４ｓｍ（２ｓａｔ）＋４ｓｔ或２ｎ＝３４＋２Ｂ＝２６ｍ＋４ｓｍ（２ｓａｔ）＋４ｓｔ＋２Ｂ。泡沙参Ａ．ｐｏｔａｎｉｎｉＫｏｒｓｈ的核型公式为２ｎ＝３４＝２８ｍ（２ｓａｔ）＋４ｓｍ＋２ｓｔ。它们同以往报道的其它种的核型相似：以中部着丝点染色体（ｍ）为主,至少具一对近端着丝点染色体（ｓｔ）和一对随体染色体,核型的对称程度较高,着丝点端化值（Ｔ．Ｃ）为５８．４％～６２．０％。结合其它性状,讨论了裂叶沙参的特殊性     

青海南部太白韭4居群的核型研究

研究了葱属太白韭青海４个居群的染色体数目和核型。结果如下,居群１：２ｎ＝２ｘ＝１６＝１２ｍ＋２ｓｍ＋２ｓｔ（２ＳＡＴ）,居群２：２ｎ＝２ｘ＝１６＝１４ｍ＋２ｓｔ（２ＳＡＴ）;居群３：２ｎ＝４ｘ＝３２＝２４ｍ＋４ｓｍ＋４ｓｔ（４ＳＡＴ）,居群４：２ｎ＝２ｘ＝１６＝１４ｍ＋２ｓｔ（２ＳＡＴ）＋Ｂｓ（０－２）。并讨论了多倍体和Ｂ染色体形成与分析。     

红江橙体胚发生及影响因素的研究

红江橙花后４周的幼果,直径约１ｃｍ左右,这时期的胚珠最适于胚性愈伤组织的诱导。以ＭＴ为基本培养基,加上适当浓度的ＩＡＡ、ＢＡ和ＭＥ可显著提高胚性愈伤组织诱导率。体胚发生,以ＭＴ＋ＩＡＡ０．１ｍｇ１－１＋ＢＡ（ＺＴ）１ｍｇ１－１培养基较好,能诱导产生正常体胚,且频率较高。胚性愈伤组织继代次数对体胚发生也有一定的影响,继代６次以内,胚性愈伤组织具有旺盛产生体胚能力;继代至第９次时,产生体胚的能力明显下降;至第１２代时,不能产生胚状体。成熟胚转换成小植株,以ＭＴ＋ＧＡ２ｍｇ１－１＋ＮＡＡ０．１ｍｇ１－１培养基成苗率最高。     

Observation on Differentiation Potential and Chromosome Stability of Callus in Subcultures of Picea wilsonii

Embryogenic calli of Picea wilsonii Mast. cultured on modified 59 medium supplemented with 1 ppm 2, 4-D and 1 ppm Kt still remained high reproductive potential after sequential monthly subcultures for 3 years. When the callus were transferred to 1/2 strength modification 59 medium with 1 ppm ABA, a lot of somatic embryos were produced after culturing about 3 monthes. The frequency of callus embryogenesis was above 90 %. Chromosome counting showed that the chromosome ploidy of callus cells of P. wilsonii was stable in subcultures for 3 years. The chromosome number and karyotype formula is 2n= 24= 16m (6sc) +8sm+2B. This result agrees with that form root-tip squash of seedling.     

Study on the Hybrid Endosperm Culture of Wheat-Rye In Vitro

The endosperm culture of wheat-rye hybrid was studied in order to explore a new pathway of chromosome engineering. The preliminary results were obtained to show that the endosperm callus formation could be induced from the young endosperm within 7–14 days after crossing on the medium supplemented with 2 ppm 2,4-D, 0.5 ppm kinetin and 3%–8% sucrose. The induction frequency of callus amounts to 35.3%. When the calli were transfered onto an auxin step-down medium containing 0.5 ppm IAA and 1 ppm kinetin, both shoots and roots were formed. 4 endosperm plantlets were obtained. The chromosome number in somatic cells of endosperm plantlets was very unstable. The numbers varied from 6—42, but there is no 49 to be found. The chromosome number with 1—4 times of 7 can be found in higher percentage.     

Induction of Endosperm Calluses and Regeneration of Endosperm Plantlets of Asparagus officinalis

The endosperm callus has been induced from the young endosperm of Asparagus officinalis L. on the MS supplemented with auxin. The induction frequency of callus amounts to 65.9%–83.1%. When the callus was transferred to the medium supplemented with lower concentration of NAA 0.1 ppm or containing BA 1 ppm and NAA 0.5 ppm, the differentiation of shoots, roots and a few embryoids began to occur. A few calluses and embryoids can develop into plantlets. The chromosome number in the cells from the same root tip and shoot apex of endosperm plantlet is very unstable. They can be euploids (n=10, 2n=20, 3n=30, 4n=40). or aneupl0ids (n=6, 7, 17, 25, 53).     

GROWTH STUDIES OF THREE CHLOROPHYLLOUS CALLUS PHENOTYPES OF GLYCINE MAX

The establishment of three chlorophyllous callus phenotypes, Glycine max (L.) Merrill, cultured on a modified Miller's medium is described. Experiments were designed to determine the hormonal requirement necessary to maintain an adequate callus growth rate that would allow for the phenotypical accumulation of chlorophyll in all phenotypes. Addition of α-naphthaleneacetic acid and kinetin, both at 1 mg/liter, to the basal medium fulfilled this requirement. However, callus growth for these phenotypes required only an exogenous supply of cytokinin. All callus phenotypes, when maintained on 3% sucrose, were shown to possess similar growth curves; however, optimal growth rates of these cultured phenotypes occurred on different levels of exogenous sucrose (NG, 2%; LG, 1%; Y, 2%). Sucrose (filtered and autoclaved) and, in most cases, fructose (filtered), when employed as a carbon source in the basal medium, maintained adequate growth rates. Glucose (filtered) supported only minimal callus growth. These callus phenotypes, after two years in culture, showed a certain degree of cell type differentiation as indicated by the formation of isolated tracheoids and in some cases organized tracheoid development. The chromosome complement (2n = 40) was observed to be polyploid.     

银鹊树胚性愈伤组织继代培养过程中的细胞染色体数目变异

以银鹊树幼嫩的合子胚为外植体诱导胚性愈伤组织,胚性愈伤经增殖后转接到液体培养基中悬浮继代培养,对其多次继代培养的胚性愈伤细胞的染色体数目检测发现:多次继代培养后的胚性愈伤细胞染色体数正常的比例为48.28%(2n=30),部分细胞出现染色体数目2n=15~60的变异,其变异率高达51.72%,其中以亚二倍体变异为主(40.07%).结果表明,在体细胞胚胎诱导形成过程中,胚性愈伤组织细胞在染色体水平上发生部分变异,这可能是体胚形成过程中畸形胚产生的根本原因.     

Studies on the Anther Culture of Triticale

The present paper deals with the study of factors affecting the frequency of induction of Triticale pollen callus in anther culture. The following experimental results have been obtained: 1. The N6 medium and B5 medium were superior to MS medium in the production of pollen callus, and the difference between N6 medium and B5 medium was not obvious. 2. When 6 to 12 per cent of sucrose was added in medium, the induction frequency of pollen callus was higher, however, this higher frequency decreased with the concentration of sucrose down to 3 per cent. 3. It was found that there was no regultr correlation between the frequency of the formation of the pollen callus and concentrations of 2,4-D (0.5 ppm, 2 ppm, 5 ppm and 10 ppm) in medium. 4. Induction frequency of pollen callus increased apparently when active carbon (0.5 per cent) was added. 5. The liquid medium was capable of improving yields of the pollen callus. 6. It is not important for the formation of the pollen callus whether the cultures were kept in light or in dark. 7. Before inoculation the spikes were inserted into a bottle with N6 medium solution and pretreated at 3–5℃ for 3 days, the frequency of callus formation may be efficiently increased in comparison with those treated in water. When N6 medium was supplemented with 2,4-D (2 ppm), NAA (2 ppm) and kinetin (0.5 ppm), either used singly or in combination, the frequency of callus formation was not obviously increased.     

Plant regeneration from immature embryos of 48 elite CIMMYT bread wheats

Forty-eight bread wheat (Triticum aestivum L.) released cultivars and elite advanced lines were evaluated for their ability to produce embryogenic callus using three different media. Basal N6 medium supplemented with dicamba (E1), MS medium containing 2,4-D (E3) or MS medium containing 2,4-D plus different amino acids (E5) were used for callus initiation and maintenance. Plant regeneration was achieved on basal MS medium with indole-3-acetic acid (IAA) and 6-benzylamino purine (BAP) and rooting on MS with 1-naphthaleneacetic acid (NAA). Percentage regeneration varied widely with both genotype and initiation medium, with values ranging from 2% to 94%. The number of plantlets produced per embryo ranged from 6 to 42. Thirteen genotypes showed at least 50% regeneration after culture on E5 medium; 3 genotypes after culture on E3 initiation medium and 1 after initiation on E1. After four subcultures, over a 16-week period, 41 genotypes (85%) lost their ability to regenerate plants while the remaining 7 lines (15%) retained plant regeneration potential but at reduced levels. E3 medium was found to be the best for maintaining regeneration potential after four subcultures.     

黄槐叶片培养过程中器官发生的研究

以黄槐（ＣａｓｓｉａｓｕｒａｔｔｅｎｓｉｓＢｕｒｍ．ｆ．）幼嫩叶片为材料,接种于ＭＳ＋ＮＡＡ１ｐｐｍ＋２,４－Ｄ１ｐｐｍ＋６－ＢＡ２ｐｐｍ的培养基上,诱导形成两种形态的愈伤组织,即致密愈伤组织与雪花状愈伤组织．将愈伤组织转移到ＭＳ＋ＮＡＡ：１ｐｐｍ＋６－ＢＡ２ｐｐｍ的分化培养基上．仅致密型愈伤组织经过球状体至不定芽途径形成大量再生植株。扫描电镜及组织细胞学观察表明,致密愈伤组织表层细胞排列紧密,有许多分生细胞团,而雪花状愈伤组织表层细胞薄壁化,分裂能力很低。球状体起源于致密愈伤组织表层的分生细胞团,其细胞有极强的分生能力,顶端可以分化发育成不定芽原基,最后形成不定芽并发育成小植株。球状体可以看成是具有形成不定芽能力的繁殖单位．     

Leaf callus induction and suspension culture establishment in lychee ( Litchi chinensis Sonn.) cv. Huaizhi

This study reports a protocol for leaf callus induction and suspension culture establishment in lychee cv. Huaizhi. The results showed that 12-day-old leaf explants cultured under a photoperiod of 16/8 h with their adaxial side touching the medium were the optimum conditions for leaf callus induction. Globular embryos were formed when the induced calli were kept on the callus induction medium without 2,4-D for 24 weeks. Friable calli were induced after 2–3 subcultures at 4 weeks intervals on the Murashige and Skoog medium supplemented with 3 mg/L IAA and 2 mg/L BAP. Suspension culture was established when these friable calli were subcultured six times in liquid callus induction medium.