Microspore culture was performed as

Microspore culture was performed as previously described by Lu et al. (2008). Briefly, the collected spikes were subjected to cold pretreatment at 4 C for 2 weeks, and the microspores were collected by crushing the anthers from the sterilized spikes. The collected microspores were isolated by filtration and centrifugation, and finally cultured on the induction medium (N6 basal medium supplemented with 2.0 lM 2.4-D, 2.3 lM KT, and 0.25 M maltose) for embryogenic callus induction. The isolated microspores cultured on the induction medium were used as control group (Control group), and those cultured in the medium containing 500 mg L -1 NaCl were designed as salt-treated group (Salt group). Four biological replicates were prepared for each groups. The microspores in each biological replcate were isolated from at least 20 donor plants, and equally divided into two parts used for two groups. After 21 days of culturing, the formed embryogenic calli in each dish (one replicate) were separated from liquid culture medium and measured to calculate the callus yield per dish. Small amount (*30 mg) of embryogenic calli were collected into tubes by snap frozen in liquid nitrogen and then stored at-80 C for RNA isolation. The remaining embryogenic calli were weighed again and transferred to differentiation medium(MS basal medium supplemented with 2.2 lM 6-BA,7.0 lM KT, 0.25 lM NAA and 83.3 mM maltose) as described by Lu et al. (2008) for plant regeneration (without salt treatment). The numbers of green plants per dish were
counted after 1 month of culturing

結果 (中文) 1:

小孢子培养技术进行如上文所述的路等人 （2008 年）。简单地说，收集的穗状花序遭受到 2 个星期，在 4 ℃ 低温预处理和小孢子被收集的破碎消毒的穗状花序从花药。收集小孢子分离过滤和离心，和最后的胚性愈伤组织诱导培养基上诱导培养基 （辅以 2.0 的 lM 2.4-D、 2.3 lM KT 和 0.25 M 麦芽糖 N6 培养基）。小孢子诱导培养基上培养被用作控制组 （对照组），和那些在含有 500 毫克 L-1 培养基培养氯化钠被设计成盐治疗组 （盐）。为每个组编写了四个生物复制。在每个生物的 replcate 小孢子分离从捐助者至少 20 家工厂，并同样分为两个部分用于两组。经过 21 天的培养，形成胚性愈伤组织在每个盘子里 （一个复制） 从液体培养基中，分离和测定，计算每道菜的愈伤组织产量。小金额 (* 30 毫克) 的胚性愈伤组织被集中到管由管理单元在液态氮中冷冻，然后储存在 80 C rna。其余的胚性愈伤组织被再次称重和转入分化培养基 (MS 基本培养基上 2.2 lM 6 — ba，7.0 lM KT、 0.25 lM NAA 和 83.3 毫米麦芽糖) 所述由路等人 （2008 年） 为植株再生 （无盐处理）。绿色的植物，每道菜数1 个月的培养后计数

結果 (中文) 3:

小孢子培养为鲁等人先前所描述的进行。（2008）。简而言之，收集的尖峰进行低温预处理在4℃，2周，和被粉碎的花药小孢子从消毒后的尖峰收集。通过过滤和离心分离收集的小孢子，最终培养的诱导培养基（N6培养基中添加2流明、2.3流明，KT和0.25米的麦芽糖）的胚性愈伤组织的诱导。的游离小孢子培养的诱导培养基作为对照组（对照组），并培养在培养基中含有500 mg L - 1的NaCl为盐治疗组（对照组）。为每一组制备了四个生物复制。从至少20个供体植物中分离出的每个生物replcate小孢子，分为两部分用于两组。经过21天的培养，在每一个菜（一个复制）形成的胚性愈伤组织分离培养液培养基和测量，计算每盘愈伤组织的产量。少量（30毫克）胚性愈伤组织收集到管快速冷冻在液氮，然后储存at-80 C RNA的分离。剩下的胚性愈伤组织再分化培养基重转（MS基本培养基附加6-BA 2.2流明，7流明0.25流明的KT，NAA和83.3毫米的麦芽糖）由路等人描述。（2008）用于植物再生（不含盐处理）。每盘菜的数量是培养1个月后计数