ESSE 2017

Proceedings of the International Conference on Environmental Science and Sustainable Energy Ed.by ZhaoYang Dong

Yong Wang

• Publisher: De Gruyter
• Year: 2017
• Print ISBN:
  9783110539134
• eBook ISBN:
  9783110539141
• Topic: Physical Sciences
• Subtopic: Environmental Science

Environmental science is an interdisciplinary academic field that integrates physical-, biological-, and information sciences to study and solve environmental problems. ESSE - The International Conference on Environmental Science and Sustainable Energy provides a platform for experts, professionals, and researchers to share updated information and stimulate the communication with each other. In 2017 it was held in Suzhou, China June 23-25, 2017.

Xia-Fei Duan, Xiao-Mu Chen, Jian-Peng Lv, Ou-Jing Li, Jin-Wang Li, Ling Wang and Zhong-You Pei*

Study on Immature Embryos of Sorghum Tissue Culture

Xia-Fei Duan, Agriculture Resourde and Environment College, Tianjin Agricultural University, Tianjin, China. Email: [email protected]

Xiao-Mu Chen, Agriculture Resourde and Environment College, Tianjin Agricultural University, Tianjin, China. Email: [email protected]

Abstract: The callus induction and differentiation of sorghum were studied in three different treatments. The results showed that the callus induction rate of 2,4–D and ZT was different when the concentration of 2,4–D was 1.0 mg/L and the concentration of ZT was 0.1 mg/L and the genotype of Xin Liang 52 had the highest induction rate of 59.3 %. The callus differentiation rate of three genotypes was different. Among them, 07–27 differentiation rates were the highest, 16.2 %. Followed by Xin Liang 52, 13.1 %. The differentiation rate of M81-E was the worst, 10.3 %.

Keywords: Sorghum, ZT, immature embryos, 2, 4–D

1Introduction

Sorghum is an annual herb of sorghum. It is a very important crop which is the world’s sixth largest crop [1]. Sorghum, a drought-tolerant crop that grows on barren land where other crops cannot grow, is widely cultivated globally. Besides, sorghum is a major contributor to some 50 billion people in semi-arid tropics of Africa and Asia, more than 30 countries Diet [2]. Sorghum can be used as human food and animal feed, but also can be used as raw materials for industrial alcohol production and bio-energy [2].

Sorghum tissue culture is late in development of plant tissue, the earliest sorghum explant culture research is Strogonol in 1968 using the root and section, callus was induced in the additional 2,4-D l mg/L, KT l mg/L MS medium, and the callus were compared with callus of Salicornia europaea L, Melilotus suaveolens Ledeb, and Brassica oleracea L.. So, the history of sorghum tissue culture research has been unveiled. Plant tissue culture provides the appropriate recipient cells for gene transfer, and also provides favorable conditions for plant regeneration and trait performance. It is helpful to solve the problems of low genetic variability, low heritability and long time for breed improvement in sorghum breeding research [3]. Compared with other crops, sorghum is difficult to obtain plant regeneration through tissue culture. The reason is mainly high dependence on genotypes and phenolic compounds [Shi Taiyuan 2003]. First of all, studies have shown that in the process of tissue culture, callus induction rate and differentiation rates of different genotypes are significantly different. So the genotype of sorghum tissue culture is very important [4]. Secondly, phenolic compounds can inhibit the callus of sorghum induction and differentiation, thus reducing callus induction rate and differentiation rate. Through continuous optimization of sorghum regeneration system, sorghum tissue culture has also made some progress. The regenerated plants were obtained from in vitro culture of immature embryos, anthers, young spikes and young leaves. Although the regenerated plants were obtained from the above explants, the induction and regeneration rates of sorghum tissue culture were very different. And it dependent on the genotype. Therefore, it is necessary to study and identify genotypes, and further improve and optimize sorghum regeneration system. Experiments of Zhao et al. (2009) showed that M81-E is a better genotype in mature embryo culture. In this experiment, we used XL52, 07–27, M81-E as experimental materials to study the induction and differentiation on immature embryo of sorghum.

2Materials and Methods

2.1Materials

Immature embryos of sorghum: XL52, 07–27, M81-E were supplied by Crop Genetics Key Laboratory of Tianjin Agricultural, planted in the crop specimen garden of Tianjin Agricultural University. Young seeds are pollinated 12–15 days later. The young seed surface was wiped with 75 % ethanol and then packaged in plastic wrap and placed at 4 °C for 12–24 h.

2.2Methods

2.2.1Medium

(1)Induction medium of sorghum callus

(2)Subculture medium of sorghum callus: MS +1.5 mg/L 2, 4-D, pH 5.8.

(3)Differentiation medium of sorghum callus: MS + 0.1 mg/L ABA + 0.25 mg/L NAA + 0.1 mg/L 1.0 mg/L ZT + 8 g/L D-Sor + 30.0 mg/L hygromycin + 1000.0 mg/L Cef + 50.0 mg/L Amp, pH 5.8

Tab. 1: Induction medium of sorghum callus

2.2.2Callus Induction of Immature Embryo

The pre-treated young seeds were carefully removed, washed with distilled water for 1 times, and then sterilized with 75 % alcohol for 1 minute, and then sterilized with HgCl2 (0.1 %) for 20 minutes, and finally rinsed with distilled water for 7–10 times. The treated seeds were placed on sterile filter paper and allowed to dry. The young embryos of the seeds were extruded in the clean bench and inoculated on the configured induction medium. Finally, the cultures in a constant temperature incubator at 28 ± 1 °C for 10 to 14 days in the dark, and then to obtain statistical induction rates.

2.2.3Sorghum Callus Subculture

After 10-14 days of induction, callus was formed on the small scutellum of immature embryo and the callus was inoculated into subculture medium with tweezers. And then the cultures in a constant temperature incubator at 28 ± 1 °C for 2 weeks in the dark.

2.2.4Sorghum Callus Differentiation, Rooting and Transplanting

The callus of good growth was inoculated on the differentiation medium, the cultures in a constant temperature incubator at 25 ± 1 °C, the daily light 16h, light intensity 2000LUX. After 4 to 5 weeks of differentiation, the callus differentiation rate (3 cm height) was calculated. When the regenerated shoots grow to about 5 cm, they are transfered to the rooting medium. When the root of the regenerated seedlings is strong, the plants are transplanted into pots containing vermiculite and nutritive soils, and until the plants mature in the greenhouse.

2.2.5Statistical Analysis of the Data

Callus induction rate = (callus number / number of immature embryos) × 100 %;

Callus differentiation rate = (the number of regenerated shoots / the number of callus inoculation) × 100 %.

After the data were transformed, the variance analysis was performed with SPSS statistical software, and the significance level was analyzed with Duncan’s new multiple range method.

3Results and Analysis

3.1Callus Induction of Immature Embryo

The results showed, the morphological characteristics of callus were different in different genotypes. The callus induced by XL 52 immature embryo was white and the ...

Table of contents

• Cover
• Title page
• Copyright
• Preface
• Committees
• Keynote Speech I Decentralized SIEPON-Based ONU-Initiated Tx/TRx Energy-Efficiency Mechanism in EPON
• Keynote Speech II A Glass Half Full or Half Empty: Water Resouces in Western China – an Earth Observation and Data Analytics Perspective
• Keynote Speech III Heterogeneous Catalysis and Environmental Nanomaterials for Water Decontamination: Promises, Challenges and Research Needs
• Keynote Speech IV Computational Studies of Molecules and Materials Pertaining to Environmental Science and Sustainable Energy
• Table of Content
• Session 1 Environmental Science and Ecology
• Session 2 Strategy of Sustainable Development
• Session 3 Energy Science and Technology
• Session 4 Control Theory and Application
• Session 5 Materials
• Session 6 Artificial Intelligence and Computer Science
• Session 7 Sensors
• Session 8 Control System and Automation
• Session 9 Advanced Design and Manufacturing
• Session 10 Emerging Fields and Other Related Fields