Shulou(Shulou.com)11/24 Report--

How many people live on the earth at this moment? Population clock will tell you that the global population has exceeded 8 billion. Among them, 702 million to 828 million people are in a state of hunger. In 2055-2058, the global population will exceed 10 billion, and by then, food will still be a big problem.

In the face of such a large population, how to "have grain in the bowl and not panic in the heart"? Staple food crops are indispensable, such as corn, the crop with the highest acreage and yield in the world. However, according to the current climate change trend, by the end of the 21st century, climate change will lead to a reduction in the area suitable for maize growth, and the average yield will fall by 6% to 24% [2]. Therefore, new technology must be used to improve maize varieties in order to meet the needs of mankind in the future of continuous warming.

Gene editing is probably one of the most promising breeding techniques in this century. although it has been used for less than 10 years, it has attracted the attention of governments, scientific research institutions and capital in recent years. In this article, let's take a look at the advantages and disadvantages of this new technology, how countries regulate, what experiences can be used for reference and what problems need to be solved urgently for China, which urgently needs to revitalize the seed industry and ensure food security.

Li Tuo, Wu Yunfei, author

Li Tuo (Editor)

Since 2021, the voice of "seed industry revitalization" is getting louder and louder. In 2023, "seed industry revitalization" has been named [3] [4] [5] by the Central Committee, the first document of the Ministry of Agriculture and villages, and the government work report during the "two sessions". It has become a hot topic, and the tone of "speeding up the industrialization of corn and soybean biological breeding" in the document is also concerned by the industry and capital.

However, from the point of view of yield and technology, there is obviously a problem of "big but not strong" in the R & D and cultivation of maize varieties in China.

1. Chinese corn, which is large but not strong, is the crop with the largest planting area and the highest total yield in the world.

China is a big country in corn planting and production. Since 2000, the planting area of corn in China has gradually exceeded that of wheat and rice, becoming the largest crop in the country. In 202112022, the global corn output reached 1.16 billion tons [6], the highest in the United States, reaching 383 million tons, and China ranked second in the world, reaching 277 million tons [6] [7].

China is also a big importer of corn. In 2022, China spent more than US $7.1 billion on 20.62 million tons of corn from overseas [8], of which the largest source of imports was the United States, reaching 16.1 million tons / US $5.26 billion [9], followed by Brazil with a total of 1.165 million tons [10]. Behind the huge amount of imports is the high domestic demand and the slow growth of per unit yield.

How big is the gap in corn yield between China and the United States? The following picture shows the comparison of the unit yield of the five largest corn producing countries in the world.

From 2000 to 2021, the top five countries in global corn yield per hectare (hg / ha, 100g / ha), the yellow line represents the United States, and the green line represents China. Why is the per unit yield of corn in China not high enough by the Food and Agriculture Organization of the United Nations (FAOSATA)? Climate, water and fertilizer, technology and other conditions are important factors affecting maize yield, and seeds are the top priority. In order to improve the quality of maize seeds, we must rely on the technical level of the breeding industry. Rong Tingzhao, academician of the Chinese Academy of Engineering, expert in maize genetics and breeding, and honorary director of the Maize Research Institute of Sichuan Agricultural University, pointed out that the shortcomings of China's corn seed industry are reflected in [11]:

Seed resources are insufficient, most of them rely on introduction, most of the existing new varieties are imitated breeding or modification improvement, and there are few independent original breakthroughs; there is also less basic research on germplasm resources, so it is difficult for theory to guide practice.

At present, the breeding technology is in the stage of fusion to "Molecular breeding 3.0", and the techniques of gene editing and artificial intelligence breeding are still in the stage of research and development.

The efficiency of industrial R & D organization model is not high, and the low-level scientific research is scattered and repeated.

The experience of the United States, Brazil, Argentina and other American countries shows that the use of high-quality provenances, such as genetically modified maize seeds, combined with special plant protection products, can significantly increase the yield. In fact, China was one of the first countries to plant genetically modified crops (the planting of genetically modified cotton was approved in 1998), and the planting area of genetically modified crops once ranked second in the world [12], but after 2010, the domestic policy attitude towards genetically modified corn breeding technology tends to be cautious, and the commercialization of genetically modified corn in China has experienced a "lost decade".

In the past two years, transgenic technology has been paid more attention to. In 2022, the Ministry of Agriculture and Village issued two batches of approval list of Safety certificates for Agricultural genetically modified organisms in 2022 [13] [14]. A total of 11 genetically modified maize varieties in China have obtained biosafety certificates. At present, many small and medium-sized breeding companies have reached cooperation with transgenic giants to introduce their own backbone inbred lines into some versions of transgenic traits.

However, there is still a lot of gap between China and foreign countries in R & D technology and policy management of genetically modified corn. With the rise of gene editing and breeding technology, domestic scientific research institutions and seed companies, transgenic and gene editing of new opportunities should not be missed, especially gene editing technology, pull up the starting time of various countries, for China, the prospect is promising.

2. Gene editing breeding: the next hot spot in 2019, Nature: biotechnology (Nature Biotechnology) magazine published "crops for 10 billion people" [15] (Breeding crops to feed 10 billion). The author listed a number of advanced breeding techniques and placed high hopes on gene editing technology.

In the past few years, gene editing breeding technology is not only a prominent study of global laboratory research, but also on the eve of large-scale commercialization, and has become a new highland of breeding technology competed by many countries.

What are the advantages of gene editing and breeding? The so-called gene editing breeding refers to the process of modifying the target genes of crops (mainly knockout of genes, gene insertion and replacement of corresponding traits of varieties, etc.), artificially creating variation, and then producing new varieties. It is the most representative 4.0 breeding technique after wild domestication, hybridization and transgenic.

The genes involved in gene editing technology mainly come from different chromosomes or varieties. For knockout gene editing, foreign genes will not be introduced. Crops bred by this method can be treated in the same way as natural variation or artificially selected varieties, while insertion and replacement gene editing may be treated as transgenic technology.

There are many tools for gene editing breeding, including zinc finger nuclease (zinc-finger nucleases,ZFNs), transcriptional activator-like effect nuclease (Transcription activator-like effector nuclease,TALENs) and protein system related to regularly spaced clusters of short palindromic repeats (Clustered Regulatory Interspaced Short Palindromic Repeat / CRISPR-associated systems, namely CRISPR / Cas).

Among them, the disadvantage of ZFNs is that the ability to produce high-frequency mutation is limited, while the disadvantage of TALENs is that it is difficult to avoid off-target effect, easy to produce harmful mutation and difficult to produce ideal mutation traits. On the other hand, CRISPR / Cas tool can effectively avoid the defects of both, and can edit genes efficiently and accurately, so it is the most commonly used technology in gene editing and breeding. [16]

The following table shows the milestones in maize breeding technology since the 1930s.

Comparison of the history and details of three technologies in maize breeding, source reference [16], tabulation, new technology of hard fruit shell naturally has new advantages: compared with artificial domestication and cross breeding, gene editing technology can achieve accurate operation, backcross takes less time, and can greatly improve breeding efficiency; compared with transgenic technology, gene editing is more accepted by the public and global supervision, and the approval process is faster. Compared with previous gene editing techniques such as zinc finger, CRISPR / Cas technology has the advantages of simple operation, high efficiency and low cost.

The general process of gene editing of plant cells using CRISPR / Cas technology, source: reference [17], redraw: shell hard science and technology humans have edited these maize genes since about 10,000 years ago, humans have domesticated ruminant grass (teosinte) and obtained corn (maize), which is widely cultivated today. Modern maize has 2.4 billion base pairs, about 30,000 ~ 40,000 genes [16].

It is generally believed that artificial domestication takes at least 20 generations to change the phenotype of wild materials, but because only a few hundred genes have changed in the long history of domesticating corn, some researchers have pointed out that with gene editing technology, even if you start "re-domestication" with ruminant grass, you can edit less than 100 genes and complete the results of thousands of years of human domestication in a short period of time. [18]

The first human use of CRISPR for gene editing of corn began in 2014 and was completed by a research team from the State key Laboratory of Plant Physiology and Biochemistry of China Agricultural University. [19]

Nowadays, by editing genes, maize can achieve the characteristics of increasing yield (by optimizing inflorescence structure, increasing photosynthesis, nutrient absorption efficiency), improving quality (nutritional value), enhancing stress resistance (herbicide resistance, drought tolerance, saline-alkali tolerance, lodging resistance), and producing male sterile lines.

By the end of 2021, researchers had edited these maize genes--

Gene editing for yield, source reference [20], tabulation (fruit shell hard science and technology)

Gene editing for quality improvement, source reference [20], tabulation (fruit shell hard science and technology)

Gene editing for stress resistance, source reference [20], tabulation (fruit shell hard science and technology)

Gene editing for male sterility, source reference [20], tabulation (fruit shell hard science and technology)

The gene editing work carried out for haploid, source reference [20], tabulation, new technology of fruit shell hard science and technology is an important factor in the development of agricultural production, but it is not the only factor. In the past 40 years, the development experience of global seed companies tells us that good technology needs the cooperation of a series of human factors in order to coruscate vitality.

Behind the high yield of corn is the blessing of seeds, while behind the seeds is the omni-directional competition of seed companies in technology, model and ecology, which may bring some inspiration for the revitalization of China's seed industry.

3. Success = technology +? In the past hundred years, European and American seed companies have experienced open competition, giant mergers and acquisitions and integration, and the development experience gained is worth using for reference.

Keep an eye on the timing and quickly follow up on new technologies. In June 1980, the United States Supreme Court ruled in Diamond v. Chacrabatti [21] (Diamond v. Chakrabarty,447 v. 303) that "whether an invention is biological has nothing to do with whether it is patentable or not." After legal guarantee, the research and development of seed companies began to speed up: in 1981, Monsanto set up a molecular biology research group [22]; in 1982, Monsanto completed its first plant genetic modification and began field experiments five years later; since the late 1980s, Monsanto has spun off chemical businesses that have nothing to do with agriculture and limited room for growth, focusing on crop production and agriculture.

Continue to invest heavily to support the research and development of new products. Domestic seed companies are often criticized as "thousands of seed companies, R & D investment is less than a Monsanto". It is no exaggeration to say that the R & D investment of multinational seed companies in Europe and the United States is generally about 10% of sales, which can be described as desperate: in 2022, Codihua's total sales are US $17.455 billion and R & D expenditure is US $1.216 billion [23] The Bayer crop Science Division (including the Monsanto business) has total sales of 25.169 billion euros, R & D expenditure of 2.876 billion euros and R & D staff of 7700, releasing 500 new varieties and hybrids within a year [24].

Pay attention to the construction of new revenue model. Most foreign agricultural giants have bundled genetically modified seeds with special pesticides to form a "no-tillage agriculture" model of "herbicide + herbicide-tolerant seeds", and "herbicide tolerance" has become a necessary character of seeds in the transgenic era. Kodihua (including its predecessor DuPont Pioneer / Dow Yinong) and Monsanto's genetically modified seeds and pesticides have achieved mutual success.

Monsanto also explored the authorization model. At first, Monsanto tried to be a "solution provider". In the early 1990s, Robb Fraley, then deputy director of research at Monsanto's department of agriculture, envisioned that it should become the Microsoft of agriculture by selling genes like Microsoft sold its operating system. Therefore, in 1992, Monsanto sold high-quality transgenic traits such as insect resistance gene (Bt) and herbicide resistance gene (HT) to rival pioneer seed industry through "one-time license" at a low price, of which Bt sold for US $38 million and HT gene for only US $500000. However, due to doubts and pressure from the company's senior management on this model, Monsanto changed its licensing model from its cooperation with Dai Zi Mian in 1993: it charged a percentage fee in the price difference between genetically modified and traditional seeds; in 1996, it was iterated into "gene use fee" (also known as "trait license fee"). Monsanto divides the seed price into two parts: seed fee and gene use fee. [25] [26]

In markets such as Argentina and Brazil in South America, "gene royalties" contribute high profits to Monsanto [27]. In order to ensure the sustainability of the market for genetically modified seeds, Monsanto will ask buyers to sign an agreement to guarantee that they will not keep seeds privately. Once found, Monsanto will sue for compensation at the standard of US $800 per acre, and in order to encourage farmers to report each other, the company even released a hotline number that received more than 1500 reports within three years [25].

Vertical and horizontal expansion and reorganization. The global seed industry has experienced three waves of large-scale mergers and acquisitions [28]. During the first wave of mergers and acquisitions (1997-2000), agrochemical companies such as DuPont, Monsanto, Dow Chemical and other agrochemical companies mainly acquired seed companies. Novartis Agriculture merged with AstraZeneca to form Syngenta, one after another to build a "germplasm resources + seeds + special pesticides" model. During the second wave of M & A (2004-2008), the agricultural giants headed by Monsanto realized the diversity of seed varieties through horizontal mergers and acquisitions; during the third wave of M & A (2016-2018), it was mainly large-scale mergers and acquisitions and restructuring of transnational capital. Bayer took Monsanto (part of the business was spun off to BASF), and Dow and DuPont spun off Kodihua after the merger. In the recent major changes in international agricultural enterprises, the previously silent Chinese capital finally entered the world seed industry: Sinochem acquired Syngenta.

By 2018, the global seed industry will form a pattern of "two super top four": the first echelon: Bayer and Kodihua; the second echelon: Syngenta, BASF, Limaglan and Kovosch. [29]

Extend to the field of AgTech. AgTech can be understood as all the modern precision agriculture that can increase agricultural income, such as digital agriculture (data analysis, artificial intelligence), new agricultural machinery (spraying with UAV, automatic irrigation of unmanned agriculture), scientific monitoring (satellite remote sensing, UAV monitoring), etc., which is characterized by paying more attention to the application of data and the use of precision agricultural technology to add icing on the cake for the high yield of excellent seeds. Global agricultural enterprises have already started acquisitions and cooperation in this field, such as Monsanto's acquisition of climate prediction company Climate Corporation, precision sowing company Precision Planting, the establishment of Climate FieldView platform.

In short, the development of overseas seed giants basically follows the path of "science and technology-oriented, finance for use, management as the key link". [26]

In 2018, Edward S., a maize breeding expert at the United States Department of Agriculture. Edward S. Buckler put forward the concept of "breeding 4.0" [30], that is, combining gene editing with interdisciplinary technologies such as synthetic biology, genomics, bioinformatics, big data and artificial intelligence to cultivate new varieties in an intelligent, efficient and directional way. He pointed out that gene editing is the last key technology of breeding 4.0.

In this era of breeding 4.0, China's seed industry ushered in a rare international opportunity, but also faced with many challenges. Below, we analyze the opportunities of China's corn seed industry in the era of gene editing from the aspects of policy, patent, business model and so on.

4. The opportunity and new technology of corn in China means new opportunities. At present, the gene editing technology in agriculture is still in the early stage of application, and China has a timely and clear attitude in formulating policies and regulatory measures; in the field of maize gene editing and breeding, China has accumulated more patents; in terms of commercialization, horizontal and vertical industrial cooperation and integration are expected to be carried out.

Under the vision of "seed industry revitalization", China's maize seed industry is faced with opportunities for reform in many aspects, such as policy, industry-university-research and ecological development.

As mentioned before the more active policy, the seeds cultivated by gene editing technology can be treated in the same way as natural variation or artificially selected varieties. Therefore, compared with transgenic, the supervision measures of gene editing issued by the agricultural department of our country are more relaxed.

In 2022, the Ministry of Agriculture and Village issued the guidelines for Plant Safety Evaluation of Agricultural Gene Editing (for trial implementation) [31]. In order to obtain the safety certificate for production and application, it is necessary to obtain at least three generations of genetic stability data. According to the efficiency of southern breeding, it may only take 1 to 2 years for a new variety to be approved smoothly, which is the same as that of the conventional major crop variety approval test, which is much higher than the application cycle of 5 years or more for transgenic safety certificate. [32]

"generally speaking, the regulation is carried out according to the GM process, and the intensity of supervision is flexible and will not be as strict as GM." Lu Hong, researcher of Shenzhen Genomics Institute of Chinese Academy of Agricultural Sciences and founder of Zhongyujin Marker and Fine Food Health Technology, told fruit shell hard technology, "there may be case by case in the process of implementation."

Looking at the world, what is the regulatory standard of China? At the end of 2022, the German scholar Torben Spendrick (Thorben Sprink) and others synthesized the gene editing supervision programs of many countries around the world [33]. According to the International access to Agricultural Biotechnology Application Service (ISAAA,International Service for the Acquisition of Agri-biotech Applications), the gene editing supervision policies of various countries are divided into two categories and four small directions [34].

Source reference [33], tabulated fruit shell hard technology Thorben Sprink pointed out that most of the countries that have issued regulatory rules (including China) belong to the "middle route" (middle ground, that is, using method 2 and method 3). The author believes that these countries take the middle path because they have high hopes for gene editing technology.

However, the core patent of CRISPR / Cas9 is not in China, and the authorization situation is crisscrossed because of the scramble for the right of patent invention in foreign countries.

The core patent staggered by canine teeth first looks at the original patent of CRISPR / Cas9.

Wisdom Bud data show that by the end of March 2023, more than 15000 CRISPR patents have been made public worldwide. The original patent distribution of CRISPR / Cas 9 is the Boulder Institute (The Broad Institute) on one side, and the team of institutions and individuals such as the University of California, the University of Vienna and Charpentier (CVC) on the other. Some patents are scattered in the Institute of Wageningen University in the Netherlands (Wageningen University and Research in the Netherlands), Toolgen in Korea, Merck in Germany (acquired by MilliporeSigma), Cellectis in France, Vilnius University in Lithuania, Kodihua and so on. [35]

Here is the global CRISPR patent application Top20 organization:

By the end of March 2023, global institutions' CRISPR patent applications, data, wisdom bud, mapping, fruit shell hard technology, whether Baird Institute, CVC, or other academic / commercial institutions, have given the green light to the CRISPR research of non-profit academic institutions, without special written authorization, but the authorization model of commercial breeding is relatively cumbersome.

First of all, you need to obtain the authorization of the Baird Institute, whose authorization model is as follows:

The patent authorization model of the Baird Institute, the source, the official website of the Baird Institute, the tabulation, the patents of the Baird Institute, which can only edit eukaryotic genes, and are not basic patents, so they also need to be obtained through ERS (ERS is an important patent agent of CVC, whose co-founder is Emmanuel Sharpentier / Emmanuelle Charpentier, one of the inventors of CRISPR), the University of California and other institutions. Obtain the basic patent authorization of CRISPR. [36]

Both MPEG LA and Kodihua (formerly Dow DuPont agricultural business) are trying to facilitate "one-stop licensing" through patent pools or cross-licensing. So far, Kodihua has won patents from Baird Institute, Caribo (CVC exclusive Licensing Company), ERS (CVC Patent Agency) and Vilnius University, together with the original research patents, a collection of the most agricultural gene editing patents.

For most domestic commercial gene editing and breeding companies, it is necessary to obtain the basic authorization of the above institutions before research and development. On the basis of the original patent, based on the CRISPR / Cas maize gene editing and breeding patent ranking in the world, China is on the top of the list. The following is the global distribution of patents for maize breeding based on CRISPR technology:

By the end of March 2023, the global institutions' patent applications for CRISPR maize breeding, data (wisdom bud, mapping) fruit shell hard science and technology, in short, in the field of maize gene editing and breeding using CRISPR tools, China has obvious advantages. However, since the original patent of CRISPR / Cas 9 is still not in China's hands, in order to avoid the potential risk of "choking", while maintaining the existing advantages, attention should be paid to the development of new genome editing tools (such as Cas 13m cas 14am cas 12f, etc.). [37]

5. To solve the real problem, the concentration of the seed industry in Europe and the United States is relatively high. In 2022, the market share (CR5) of the top five seed enterprises reached 51%. In China, even if there is a giant Syngenta, the CR5 of China's seed industry is only 11% [38].

However, the low concentration of China's seed industry is only superficial. In order to realize the "revitalization of the seed industry", we still need to solve these problems.

"the hope of the whole village" is not necessarily the head company. Industry research generally believes that the concentration of China's seed industry is not high enough to meet the huge commercial demand [38] However, front-line R & D people do not think so. for example, he Wei, founder of Tieling rising Sun, pointed out in an article in the "North-South Academy": large domestic breeding companies have received many resource projects, but there are many managers and few front-line researchers. The change in leadership has led to a lack of continuity in scientific research and "has not taken on what a big company should look like." [39] the current situation of the industry shows that the survival desire and flexibility of small and medium-sized seed companies make their innovation ability better, and the new varieties developed are easier to sell to large companies.

The depth of cooperation between head companies and small and medium-sized companies needs to be improved. Large and small companies have a long history of cooperation, but most of them only focus on variety licensing transactions. "many head seed companies do rely on buying varieties from small and medium-sized breeding units, and lack of long-term and stable in-house R & D (internal research and development)." According to Lu Hong, "it is very difficult to establish an R & D system like that of a large international company." This suggests that although the head company has established a simple variety acquisition partnership with small and medium-sized companies / institutions in the past, it is only a simple "transaction" relationship and a more mature ecology, which needs to be encouraged and nurtured by policies.

The ceiling of breeding policy needs to be reconstructed. Lu Hong pointed out that the relationship between domestic leading large companies and small and medium-sized companies needs to be coordinated and co-ordinated: "National breeding institutions should cooperate deeply with commercial companies and give full play to their respective advantages. The seamless connection between traditional breeding and modern breeding technology will be an effective strategy to improve the R & D capability of China's seed industry, and it will also be the way of industry-university-research integration that the country has always advocated. However, it requires a high degree of coordination among the three parties (seed companies, scientific research institutions, and the government), and requires leading talents with global vision and industry experience to carry out overall design. "

The driving force of basic R & D in the industry. Although domestic regulation is "loosening", due to insufficient intellectual property protection, large state-owned and listed companies lack the motivation to independently develop new varieties, do not establish a strong R & D system, and do not have sustained and stable investment. due to short-term performance-oriented pressure, large companies prefer to buy new varieties from small and medium-sized companies.

"Gene editing technology cannot replace other breeding techniques within 10 years, and cross breeding is still the fastest and most effective breeding method." Through the new technology policy or leading to industrial changes, Lu Hong believes that "the liberalization of GM and gene editing technology will bring an opportunity to reshuffle China's seed industry, but the intensity of the shock wave depends on the state's implementation of the seed Law and the protection of intellectual property rights."

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[20] CRISPR/Cas System: Applications and Prospects for Maize Improvement

Yilin Jiang, Kangtai Sun, and Xueli An. ACS Agricultural Science & Technology 2022 2 (2). DOI: 10.1021/acsagscitech.1c00253

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[37] CRISPR/Cas System: Applications and Prospects for Maize Improvement

Yilin Jiang, Kangtai Sun, and Xueli An, ACS Agricultural Science & Technology 2022 2 (2), 174183 DOI: 10.1021/acsagscitech.1c00253

[38] Minsheng Securities: the granary is solid but the world is safe, and if the provenance is strong, agriculture will flourish: seed industry in-depth report https://pdf.dfcfw.com/ pdf / H3_AP202211131580205880_1.pdf

[39] North and South Academy: laymen talk about the current situation of domestic breeding 2021-5-8 https://mp.weixin.qq.com/ s / 9sr5vU9fUbqmz1nVh0rfRw

This article comes from the official account of Wechat: fruit Shell hard Technology (ID:guokr233), author: Li Tuo

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