دانلود ترجمه مقاله کشت و امتزاج پروتوپلاست بین دو گونه کلزا carinata و napus

• فهرست مطالب:

چکیده
مقدمه
مواد و روش ها
مادۀ گیاهی
ترکیب و جداسازی پروتوپلاست
کشت پروتوپلاست
احیای گیاه
تجزیه و تحلیل داده ها
نتایج
احیای ریزکالوزها
احیای کالوزها
احیای گیاه
ترکیب پروتوپلاست
بحث
نتیجه گیری

• بخشی از ترجمه:

احیای ریزکالوزها و کالوزها در تمام ژنوتیپ های آزمایش شده مشاهده شد. تأثیر قوی ژنوتیپ ها براحیای هر دو نوع بافت dedifferentiated(غیر متمایز) ،بوسیلۀ آنالیز آماری تأیید شد. براسیکا کاریناتا BC-6 و BC-1 بهترین ها ژنوتیپ ها برحسب شکل گیری ریز کالوزها و کالوزهای پروتوپلاست های ایزوله شده (جدا شده) در اختیار ما قرار دادند. در عوض،مطابقگزارشات ، قابلیت احیا در ژنوتیپ های BC Dodolla براسیکا کارایناتا و BNSL-03/04 براسیکا ناپوس رضایت بخش نبود.اما تفاوت های قابل توجهی نیز بین رونوشت های بیولوژیکی در برخی ژنوتیپ های آزمایش شده کشف شد.علت این پدیده احتمالا به دلیل سطح معین غیریکنواختی و ناجوری مادۀ بیولوژیکی مورد استفاده برای رونوشت های پی در پی می باشد.
کل گیاهان دارای ریشه های کاملا رشد کرده از سه ژنوتیپ بودند؛ فقط کالوزهای دارای ساختارهای مریستمی سبز تیره توانستند پریموردیوم های ریشه و سپس شاخه را احیا کنند. بااینحال، برگ ها و ساقه های شکستنی hyperhydrated در تمام احیاشده های (regenerants) حاصله مشاهده شدند. درنتیجه، زیرکشت های تکراری در محیط MS جامد برای دستیابی به گیاهان قوی لازم است.
ترکیب کامل و صحیح غلظت های PEG و زمان تیمار ، تعداد ترکیب بین ژنوتیپ های مورد استفاده را تعیین کرد. با وجود اینکه محلول ۳۰% PEG به عنوان بهترین غلظت ارزیابی شد. اما مقدار زیادی از multifusants هایی که به صورت اضافی در کاربردهای عملی استفاده شدند به طور ویژه در ظروف پتری با تیمار PEG طولانی تر کشف شدند.

• بخشی از مقاله انگلیسی:

The genus Brassica includes a wide range of crop species with great economic value worldwide. Therefore, they attracted not only breeders using conventional methods but also those concerned with biotechnological methods. In recent years, major efforts in Brassica research have focused on utilization of tissue culture technology for crop improvement. Microspore culture and protoplast culture techniques are most frequently used for the manipulation of foreign gene to broaden genetic diversity. The techniques of protoplast culture and fusion are becoming increasingly useful accessories for breeding programs in various crop plants. The most widespread use of protoplasts is for somatic hybridization experiments either to overcome barriers in sexual crosses or to modify cytoplasmic traits by altering organelle populations (Kumar and Cocking, 1987). Somatic hybridization has been used in Brassica improvements programs for the transfer of chloroplastencoded triazine resistance into cytoplasms carring mitochondria-encoded male sterility (Pelletier et al., 1983) and for the transfer of male sterile cytoplasm from spring to winter rapeseed (Barsby et al., 1987). Numbers of studies have observed that the in vitro tissue culture and the regeneration of plants can lead to alterations in traits conditioned by the cytoplasm (Gengenbach et al., 1981; Kemble et al., 1984). As outlined above, Brassica intergeneric, interspecific and intraspecific somatic hybrids and cybrids have been obtained and used for the transfer of agronomically important nuclear and cytoplasmic traits. Plant regeneration has been increasingly realized via organogenesis and somatic embryogenesis by means of various explants. Protoplast culture improvements have been focused on various factors including genotypes, media additives and effects of enzyme treatment on viability of protoplasts at different time durations. The aim of our study was to identify Brassica carinata and Brassica napus genotypes or inbred lines that offer protoplasts with satisfactory regenerative capacity, to optimize current methods for establishing protoplast culture in selected genotypes of Brassica carinata and Brassica napus, to develop an efficient and reliable protoplast culture protocol for fusion experiments in selected Brassica carinata and Brassica napus genotypes. MATERIALS AND METHODS Plant material High productive doubled haploid (DH) lines of winter oilseed rape (Brassica napus L.) as a source of mesophyll protoplasts BN-OP-01 and BN-SL-03/04 and genotypes of Ethiopian mustard (Brassica carinata A. Braun) – DH line BC Dodolla and breeding materials BC-1 and BC-6 – as a source of hypocotyl protoplasts were used for experiments. To obtain sterile plant material for establishing protoplast cultures, seeds of above mentioned genotypes were surface sterilized in 70% ethanol for one minute, followed by immersion in 30% commercial bleach Savo (on the basis of NaClO) for twenty minutes and finally by washing procedure with sterile distilled water three times. MS medium without growth regulators was used for germination of seedlings. Seeds of genotypes intended for hypocotyl protoplasts were incubated in a thermostat at 25°C in the dark; those for mesophyll protoplasts were kept in a cultivation room and cultivated under controlled environmental conditions (25°C and 16/8h day/night photoperiod and light intensity 260 µmol/m²/s). Protoplast isolation and fusion Seven-day old hypocotyls and leaves from one-monthold in vitro plants were cut into 1mm segments and treated separately in different Petri dishes with the enzyme solution, containing 0.25% macerozyme R10 (Serva) and 1% cellulase Onozuka R 10 (Serva) in W5 salt solution (Potrykus and Shillito, 1986, Mukhopadhyay et al., 1994). Cut tissue was incubated for 18 hours in a thermostat at 25°C without shaking. Incubated mixtures were filtered separately through a 50 µm nylon mesh and transferred to 10 ml centrifuge tubes. Suspensions were centrifuged at 100 g for 5 minutes. After careful supernatant removal, 4 ml of 20% sucrose solution was mixed with the protoplast suspension and then gently covered with 2 ml W5 salt solution without disturbing the protoplast suspension. Suspensions were again centrifuged at 100g for 5 minutes.

دانلود رایگان مقاله انگلیسی

خرید ترجمه فارسی مقاله