Crop produce comes from seeds. It is important to have elite seeds for cultivation and harvesting. There are two major types of seeds in the seed market: F1 hybrid seeds and open-pollinated seeds (OP, traditionally pollinated). Farmers in developed countries plant F1 hybrid in most cases, while farmers in developing countries plant mainly OP. In fact, 60-70% of seeds planted in India and China are OP because OP is significantly cheaper. There are several reasons why the seed industry is important. First is for global food security. Based on the fact that the global population continues to increase steadily, additional productivity of 70% will be required to feed the global population by the year 2050. Second, seeds were traditionally used as food, both fresh and feed, but have now become materials for future industries of medicine, pharmaceutics, functional foods, energy, and may other applications. Third, new breeding programs based on biotechnology have changed the seed market dramatically. These programs are highly competitive and indeed play a major role, not only in the reduction of breeding time, the development of various genetic sources, the enhancement of purity and cost-saving, but also for the selection of value-added varieties. In Korea, F1 breeding began 65 years ago and the breeding programs for several vegetables and rice are in the top class worldwide. In addition, for the first time in 1999, a private seed company in Korea employed biotechnology for the purpose of crop breeding to develop platform technologies that could be utilized in the breeding practice. The major achievement so far is the development of DNA markers associated with resistance to disease, tolerance to the environment, and functional aspects. The application of genotyping has made many services possible, such as the purity control of F1 and inbred lines, variety verification, MAS (marker assisted selection), and MAB (marker assisted backcrossing). In addition, cell fusion and DH technologies have helped breeders to solve breeding limitations. There have been many cases of successful crop transformations, however, no GM varieties have been successfully commercialized in Korea. I bet this is inevitable, though. And it should be, because Korea imports lots of GM products, equivalent to $3 billion every year. More seed production and higher crop quality require new R&D strategies for breeding practices in the seed industry. Thanks to genomics information with big data and anti-GMO policies, new technologies are on the horizon, including genomic breeding, genome editing, in silico breeding and NBT (new plant breeding technology). I am going to talk more about the direction and strategy of R&D for crop breeding.

Bt gene derived from the B. thuringiensis has been used for developing GM crops, and corn, cotton and soybean producing B. thuringiensis toxins have been on the market for last 17 years or so creating a huge GM seed industry. One of the notorious pests in brassica crops is diamondback moth (DBM). In order to protect the insect plague of crops from DBM, 4-5 billion dollars have been wasted annually for applying integrated measures in worldwide. Major prevention is use of pesticides that may build the contamination level of chemicals in the ground and this practice threats the environment and ecosystem. An alternative is to develop GM brassica crops and therefore we have developed GM cabbages resistant DBM using bt gene. Lots of T0 cabbages were tested for resistance and independent GM cabbages resistant to DBM were selected. Molecular analysis was conducted to find if GM cabbage holds one copy transgene and intergenic insertion. We found an independent GM cabbage and it contained a singly copy of the transgene without disturbing the insertion site. This one called C95 line with an status of event have been self-crossed for two generation (T2). Also we are working the development of GM cabbage with different vector that contains bar gene as a selection marker. So far 17 T0 cabbages have been obtained by bar selection.

Many viruses infect cucurbits. One of the well-known symptoms is mosaic disease. Those that cause mosaic are cucumber mosaic virus (CMV), squash mosaic virus (SqMV), watermelon mosaic virus (WMV), zucchini yellow mosaic virus (ZYMV) and cucumber green mottle mosaic virus (CGMMV). WMV resistant GM squash was developed many years ago in the United States and it was on the market, but no further information was available by now pertinent to commercial aspect. Usually these viruses are not easily controlled by frequent applications of chemicals that target the insect as carriers of viruses. Therefore, it is necessary to develop commercial varieties possessing resistance against viral diseases. We have developed GM watermelon rootstocks called gongdae, using a coat protein gene of CGMMV as transgene. Those GM watermelon rootstocks showed highly resistant to CGMMV, and have been crossed to get the several BC and T generation. In order to obtain the virus resistant watermelon, watermelon lines were crossed to the selected GM watermelon rootstock. Here, we present the successful watermelon cultivars that show resistance to CGMMV. The resistance must have obtained by transferring the transgene from the GM watermelon rootstock to watermelon line

CMVP1 (cucumber mosaic virus pathotype 1) has been frequently occurring virus causing damage in pepper farms, and it is hard to control the outbreak due to lack of the genetic source resistant to this specific pathotype. Therefore, we have developed transgenic peppers tolerant of CMVP1 using a CP gene of CMVP0 pathogen. In order to fulfill the requirement of the biosafety assessment criteria, we have studied the horizontal gene flow from GM pepper to non-GM pepper by monitoring the transgene movement. If the pepper farms are located closely each other and the pollen moves from GM pepper to non-GM pepper, it would cause unintended fertilization. Therefore, a buffer zone to separate the cultivation regions is required to avoid the contamination of transgene. Previously, several data regarding the movement distance of pepper pollen were reported by judging the phenotypic change. However, no tool as a trace marker was available. The objective of this study was to assess the frequencies of gene flow from GM peppers to non-GM peppers in neighboring farms using the transgene of CP as a trace marker. The GM and non-GM peppers were cultivated in the isolated farm of Nongwoo Bio Co. (NW GM pepper field) and pepper fruits were collected from the NW GM pepper field as well as the neighboring pepper farms. The pepper seeds collected from the farms were planted and the massive PCR analysis was performed to answer the question how far the pollen of GM pepper migrates. The conclusive data based on the consecutive experiments for 6 years is that the gene flow by pollen movement did not occur in peppers that were separated each other over 30 m.

In Korea, CMV (cucumber mosaic virus) is the most frequently occurring virus with a single infection rate of 45%. However, a total occurrence of CMV by co-infection, either couple or multiple, with BBWV (broad bean wilt virus), PepMoV (pepper mottle virus) and PMMoV (pepper mild mottle virus) covers over 90% in the field cultivation of pepper. The PepMoV is transmitted by several aphid species, and it has been considered the most frequently detected potyvirus when it co-infects with CMV or PMMoV. Since F1 hybrid that resistant to PepMoV has not been developed, we have developed transgenic peppers using Agrobacterium-mediated transformation with a Hc-Pro gene of the PepMoV. A large number of T1 peppers were tested for resistance to the PepMoV, and T1 peppers tolerant of PepMoV were selected. After consequent self-crossing up to T4 generation, highly tolerant peppers to PepMoV were selected. So far, BC3F1 lines have been selected by back-crossing with 4 elite lines through a breeding program. The horticultural differences of the GM line comparing to inbred lines were investigated and no statistical significance between GM and non-GM lines was found. Based on molecular analysis, One of GM lines, 10-2, contained the transgene in the non-coding region indicating that this line would be a GM event.

Many viruses infect cucurbits. One of the well-known symptoms is mosaic disease. Those that cause mosaic are cucumber mosaic virus (CMV), squash mosaic virus (SqMV), watermelon mosaic virus (WMV), zucchini yellow mosaic virus (ZYMV) and cucumber green mottle mosaic virus (CGMMV). WMV resistant GM squash was developed many years ago in the United States and it was on the market, but no further information was available by now pertinent to commercial aspect. Usually these viruses are not easily controlled by frequent applications of chemicals that target the insect as carriers of viruses. Therefore, it is necessary to develop commercial varieties possessing resistance against viral diseases. We have developed GM watermelon rootstocks called gongdae, using a coat protein gene of CGMMV as transgene. Those GM watermelon rootstocks showed highly resistant to CGMMV, and have been crossed to get the several BC and T generation. In order to obtain the virus resistant watermelon, watermelon lines were crossed to the selected GM watermelon rootstock. Here, we present the successful watermelon cultivars that show resistance to CGMMV. The resistance must have obtained by transferring the transgene from the GM watermelon rootstock to watermelon line.

Bt gene derived from the B. thuringiensis has been used for developing GM crops, and corn, cotton and soybean producing B. thuringiensis toxins have been on the market for last 16 years or so creating a huge GMO industry. One of the notorious pests in brassica crops is diamond backmoth (DBM). In order to protect the insect plague of crops from DBM, 4-5 billion dollars have been wasted annually for applying integrated measures in worldwide. Major prevention is use of pesticides that may build the contamination level of chemicals in the ground and this practice threats the environment and ecosystem. An alternative is to develop GM brassica crops and therefore we have developed GM cabbages resistant DBM using bt gene. Lots of T0 cabbages were tested for resistance and independent GM cabbages resistant to DBM were selected. Molecular analysis was conducted to find GM cabbage to hold one copy transgene and intergenic insertion. We found two independent GM cabbages as an event and those have been self-crossed for two generation. Also we are working the development of GM cabbage with different vector that contains bar gene as a selection marker.

The most important factor in breeding program is to obtain the value-added genetic line. Generally, breeders develop genetic sources using several methods such as segregation-breeding, cross-breeding, backcross-breeding, mutation induction, tissue culture and so on. Here, we present one classical way but very valuable method called cell fusion or protoplast fusion to create genetic sources for the breeding practice. The method we developed was the asymmetric somatic-hybridization of protoplast isolated from carrots. This is rather to transfer the nucleus from the high quality F1 hybrid to other mediocre line to produce a new carrot line. Since the breeding a carrot line for higher quality and purity takes a long time, therefore this nuclear transfer technology is very beneficial to generate a new line that could be useful to breed elite varieties. We had obtained around 200 fused carrots (cybrids), 12 cybrids were self pollinated and produced seeds. Selected progenies have been evaluated for horticultural characteristics and we have found new genetic lines that show better phenotypes.